Reflector turret unit; S_M_II

Belt driven

For technicians and partly for sales managers!

 

 

These instructions describe the procedures to install and adjust the Belt driven Reflector Turret Unit (RTUB) for the scanners Pannoramic SCAN_II and Pannoramic MIDI_II. To help to resolve problems with the turret unit or problems with fluorescent scanning a functional overview, a hardware description of the used components and adjustment procedures are added.

 

The following description is based on the P250 type scanner and the slide scanner “Pannoramic SCAN_II”.

 

Precautions:  Never look directly into the beam of the fluorescent light source! The lamp emits also ultraviolet light with very high intensity. To prevent your eyes from harm (damage) use always sun glasses with a high filter factor of UV light if the fluorescent light source is switched on and you are adjusting the beam even if the cover of the turret unit is removed. For further precautions please, refer to the manual for the fluorescent light source you are using!

 

For safety regulations regarding human health and scanner functionality please refer to:   Precautions

 

 

 

 

 

Contents

 

General

Configure the belt driven reflector turret unit

Functional overview of the reflector turret unit

Construction of the fluorescent exciting and image path

Construction

Mechanical components and construction

·         Stepper motor and the belt wheel

·         Filter wheel

·         Mechanical drive

·         Diaphragm position adjustment tools

·         Iris size adjustment tools

·         Mechanical shutter

·         Remove or mount the FL reflector turret unit

 

Optical components

·         Light source adapter and mounting

·         EPI-fluorescent illumination unit

·         Mirror

·         Filter block

·         Virtual fluorescent tissue

 

Adjustment techniques / checks

·         Tools, used for the adjustments

·         Check and adjust the tightness of the belt

·         Adjust the external sensor position

·         Find the Home 1,2 position

·         Find the first filter position

·         Adjust the aperture position

·         Adjust the aperture size

·         Adjust the Luminous field position

·         Adjust the luminous field size

·         Check the correctness of the filter fixing in the filter positions

 

 Adjustment procedure

 

 

 

General

 

The reflector turret unit is a component added to the Pannoramic P250 scanner to give the possibility for fluorescent exciting and scanning of tissues; the component is not used during the brightfield scan process. For fluorescent scanning of tissues, light wave length filters are used in many variations; the filters are assembled into a filter block. The filter wheel in the turret unit has ten positions, so it can contain up to 9 filter blocks for the fluorescent scan procedure of stained tissues; in the tenth position of the filter wheel, the image cover tube have to be inserted. The inserted filters (positions) can be selected by software during the manual fluorescent scan procedure; the assigned filter(s) will be selected automatically during the automatic scan procedure.

 

 

 

The exchange of the entire turret unit is possible:

 

 

 

Requirements

·       Service program for Pannoramic scanners (SlideScanner Service.exe) with actual license file

·       Pannoramic Scan and Pannoramic Viewer software (SlideScanner.exe, SlideViewer.exe) with dongle or actual license file

·       1.5, 2.5, 3 and 5 mm hex key wrenches,

·       Hardware and construction knowledge of the Pannoramic 250.

·       Deeper knowledge of handling the Pannoramic Scan and Pannoramic VIEWER software

 

 

 

 

 

 

 

Configure the belt driven reflector turret unit

 

 

Since the software version 1.15 the units of the scanner are configured in the file “MicroscopeConfiguration.ini”, section [Microscope].

 

 

 [Microscope]

SerialNumber=xxxxx

MicroscopeType=3DMic ...

ScanCameraType=

PreviewCameraType=CVrmc_m8_pPro

BarcodeReaderType=PreviewCamera

LoaderType=SL_9Mag_25Slide_Sensor_Vertical

CameraChangerType=CC_3DH_2Pos

ReflectorTurretType=RT_3DH_10Pos_Belt

BrightfieldLightSourceType=FlashLight2010

ObjectiveChangerType=OC_2Pos

ObjectGuideXYZType=OGXYZ_FLASH3

FlashUnitType=FlashUnit_Type2

NDFilterType=NDType2

PreviewLightType=PreviewLightUnitType_Type2

PowerSwitchBoardType=PowerSwitchBoard_Type1

 

The actual version of the belt driven reflector turret unit in the scanners SCAN_II and MIDI_II is:

 

ReflectorTurretType=RT_3DH_10Pos_Belt;                        if the turret unit is implemented.

ReflectorTurretType=RT_None;                                  if the turret unit is not implemented; brightfield scan only

 

If modifications are done in the file “MicroscopeConfiguration.ini”, the scan software “SlideScanner.exe” has to be started again; only so the modifications take effect (this is true for some parts of the service program also).

 

 

 

 

 

 

 

Functional overview of the reflector turret unit

 

 

The turret unit allows the exciting of the fluorescent stained tissue via the relevant filter block and the objective.

 

The filter wheel of the turret unit has 10 positions and may so contain up to 9 filter blocks; in the 10th position the image cover tube is inserted.

 

·       The appropriate filter is selected via software buttons (or automatically) before the FL scan procedure starts and automatically if the scan process is in progress.

·       If single band filters are used, the actual filter block is often changed to a filter block with another wave length.

·       By using multi band filters, the movement of the filter wheel is seldom not required during the scan procedure and so, the scan time of the tissue is drastically reduced.

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

                   Construction of the fluorescent exciting and image path

  

The fluorescent light source is connected to the turret unit via the „Fluorescent light source adapter mounting”; the adapter itself is used to interfacing the traditional light source like the „X-Cite® Series 120” or the “Lumencor®” exciting light source. The different interfaces are realized via separate adapters. The tissue is stained and prepared to fluoresce, if it is excited with a high intensity light. The emitted light beam of the light source is prepared by the “Aperture stop diaphragm” and the “Luminous field stop diaphragm”.

The mirror reflects the light beam to the excitation filter of the “Filter block”.

If the filter block is single band and the light source emits white light, the emission filter reflects all the wavelengths of the white light, except the defined excitation wave length.

In the filter block the excitation filter, the beam splitter and the emission filter are combined for a special excitation and the relevant emission wave length.

The appropriate wave length of the excitation light beam passes thru the excitation filter and will be reflected to the objective by the help of the beam splitter.

The optics in the objective is used to illuminate the tissue and excites the used stain in the field of view.

The stain of the tissue fluoresces and the emitted light rays (in a higher wave length then the excitation wave length; with less brightness) are collected by the objective; the image passes thru the beam splitter, the emission filter, the tube lens and the camera changer to the CCD of the scan camera.

 

The wave lengths of the components (the excitation light wave length, the characteristics of the filter block and the used stain of the tissue) are combined for a specified light wavelength; this must be met by all used components, otherwise the quality of the scanned tissue is reduced or even bad.

 

 

       Optical path and Field Of View

                        Fluorescent_exciting

 

 

 

 

 

 

 

 

                   Physical solution of the exciting path

  

Traditionally, the fluorescent light beam may contain all the wavelengths from ultra violet (from about 350nm) thru the visible light (about 400nm to 780nm) until the infra red spectrum (over 780nm to 1000nm). The relevant wavelength to excite the stain (fluorophore) of the tissue is filtered and passes thru the excitation filter; all other wavelengths will be reflected. In other words, the characteristic of the excitation filter defines the light wave length to excite the stained tissue.

The filtered wavelength will be reflected to the objective by the beam splitter and so the fluorophore in the field of view of the tissue will be excited.

 

Important

The characteristic of the excitation filter and the beam splitter must meet the exciting wavelength of the fluorophore!

 

       Determining the Filter Type

 

In newer exciting light sources like the Lumencor® Spectra, the exciting light will be generated by powerful light modules which produce the exciting light wavelength directly. By switching the modules and using multi band filters, combined for more wavelengths in the same filter block (e.g. Quad Band Filters), the movement of the filter wheel can be reduced to a minimum and so the fluorescent scan procedure is less time consuming.

 

 

 

 

 

 

 

                   Construction of the belt driven FL reflector turret unit

 

1.      Exciting light source adapter mounting

2.      Exciting light source adapter fixing latch

3.      Aperture stop diaphragm

4.      Luminous field stop diaphragm

5.      Condenser

6.      Mirror

7.      Filter block

8.      Filter wheel

9.      FL image path’s cover tube

10. Belt

11. Belt leading rolls and belt tightness

12. Turret motor

13. Power and control connector

14. External Home sensor connector

15. Luminous field size adjustment

16. Aperture size adjustment.

17. Flexible shaft for the position adjustment bolt

 

 

 

 

Main modifications

            in relation to the gear driven FL reflector turret unit consist of:

 

·         The solution of the light input connection was modified; so the Lumencor® or conventional fluorescent light sources can be connected via separate, special adapters.

 

 

 

Principle of belt drive

 

If the filter wheel is in the Home1,2 position (the Home1 sensor in the stepper motor is active and the permanent magnet stays over the external sensor) the filter in the position 6 of the filter wheel can be inserted or exchanged and the filter in the position 1 of the filter wheel stays in the light and image path.

The first filter position defines the exact filter block position in the exciting / image path.

Because the external home sensor position is not always exactly on the same position (the position of the belt wheel on the motor axle in relation to Home1 and the position of the permanent magnet in relation to the external sensor), the value of the “first filter position” defines a deviation in motor steps, in relation to the found Home1,2 position.

The “first filter position” is defined in the file “MicroscopeConfiguration.ini” section [ReflectorTurret] with the parameter value of “StartingMotorPosition=”.

The deviation from the Home1,2 position is often +- some 10 steps only.

 

 

 

 

 

 

Mechanical components and construction

 

       Stepper motor

 

The stepper motor is used to rotate the filter wheel of the turret unit and so, any inserted filter (or filter position) can be selected via software commands at the appropriate moment.

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

The sensor cover and motor mounting connects the stepper motor to the turret plate.

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

                   Belt wheel

 

The belt wheel transmits the force of the rotor to the belt. The position of the belt wheel on the rotor axle is defined by the position of the belt leading rolls.

The diameter and the cog number of the belt wheel were chosen so, that the exact distance of 3200 motor steps between two filter positions is realized.

 

 

 

 

       Find the correct belt wheel position” and “Position of the belt wheel’s clamp

 

 

 

 

 

 

                   Filter wheel

 

The Filter wheel has 10 positions, so it is able to contain up to 9 filters and the fluorescent tube for the brightfield scan procedure. The filter blocks are fixed in their positions via springs. For bright field scanning and fluorescent scanning also, in the 10th filter position the image path cover tube must be inserted, or at least no filter block must be inserted in this position.

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

                         Filter springs

 

The filter fixing springs are very sensitive in question of deforming, because these springs guarantees the proper position of the filter block in relation to the optical axis.

Any improper fixing of the filter in the filter wheel (some 10th mm are important) modifies the “1st filter position” of the incorrect inserted filter block and so the straightness of the optical axis is also incorrect; this results in improper exciting of the stained tissue and reduced image scan quality of the virtual tissue; see also Check filter block positions.

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

                   Insert or remove filter blocks

 

Fit the filter block between the springs, into the filter block position of the filter wheel as shown, then press the upper edge carefully downward until the sideward pressing springs fixing the filter block correctly.

Check the proper position of the filter block in the filter wheel manually; movements on the position surface must not occur.

 

 

 

To remove the filter block, lift up the filter block carefully on the edge of the fixing springs from beside until the springs are even disconnected, then move the entire filter block sideward.

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

                   Mechanical drive

 

The belt wheel on the motor axle drives the filter wheel via the belt. The mechanical drive solution allows a filter position change with exact 3200 motor steps; in other words, the motor axle does one revolution to reach the next or previous filter position. To move the filter wheel from the first position to the 10th position the wheel goes 9 times 3200 steps forward or one filter position backward respectively.

 

The belt leading rolls define the belt position in relation to the filter wheel; because the filter wheel can be driven in both directions, two leading rolls are used.

Because the belt wheel can be moved (shifted) on the rotor axle of the motor, the adjusted belt wheel position defines the straight movement of the belt during rotation of the filter wheel.

 

Adjust the belt tightness by positioning the leading roll 1 upward or downward until no backlash can occur in the filter position and the rotation of the filter wheel and the motor is done nearly noiseless.

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

                   External sensor

 

The filter wheel is designed to rotate endless, so the motor’s internal sensor “Home2” can not be used to find the start position inside of 1 revolution of the filter wheel.

The start position of the filter wheel is defined by the implemented permanent magnet; its position is exactly between the 3rd and the 4th filter position on the filter wheel. The external Home2 sensor is implemented into the turret plate at the specified position; the sensor is externally connected to the stepper motor of the turret unit so, that the information of the motor’s internal Home2 sensor is overridden.

 

 

       Adjust the Home sensor position” and “Check or adjust the external sensor acting range

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

                   Diaphragm position adjustment tools

 

 

The adjustment bolts are used in a pair of bolts with a shorter and a longer flexible shaft and these adjust the position of the aperture and the luminous field diaphragm respectively in X- and Y-direction in relation to the light beam. The fixing bolt is also used to bearing the tool knob in the mounting. The fixing bolts for the other two tool knobs are found on the opposite side, from the top. By removing the fixing bolt the appropriate adjustment bolt can be dismounted, if necessary.

  

       Position of the diaphragm

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Handling the position adjustment tools

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

                   Iris size adjustment tool

 

With the adjustment tools “Aperture size” and “Luminous field size” the size of the iris can be defined via an ex-center. This transforms the up / down movement of the “Aperture size” or “Luminous field size” tool respectively to a rotation of the iris mechanics and this will more open or more close the iris.

Both tools can be fixed separately by screwing the tool knob.

 

If the spring was dismounted or is not fixed properly, there exists the possibility that the ex-center is disconnected from the bolt of the iris mechanics. In this case, you are unable to adjust the iris size. Check the connection between ex-center and iris bolt always after reassembling. If the open / close tool is fully pulled or fully pushed, disconnection must not occur. Check this behavior in various end positions of the adjustment bolts also!

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Handling the size adjustment tools

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

                   Mechanical shutter

 

The mechanical shutter is implemented as a part of the focus unit. The shutter must be closed during fluorescent scanning and insures a dark background. Other, unwanted fluorescent materials (e.g. painting, optics) can not reflect the fluorescent light or can not fluoresce and so they will not disturb the fluorescent view.

 

       Focus unit’, and  Shutter mechanics

 

 

 

 

 

 

 

 

 

 

 

 

 

 

                   Remove the turret unit from the scanner

 

 Remove from SCAN

1.      Remove the camera tube (if exchange).

2.      Remove the unit spacer

3.      Remove the cable FCJ-1 from the turret stepper motor.

4.      Remove or loosen respectively the mounting bolts for the turret unit as shown.

5.      Pull the entire fluorescent reflector turret unit in an angle of approximately 30 degrees upward and frontward.

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 Remove from MIDI

 

1.      Remove the camera tube (if exchange).

2.      Remove the cable FCJ-1 from the turret stepper motor.

3.      Remove or loosen respectively the mounting bolts for the turret unit as shown.

4.      Remove the unit spacer bolt, then the unit spacer and pull the entire fluorescent reflector turret unit horizontally to the right.

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

                   Insert and mount the reflector turret unit

Because the camera tube mounting is a part of the turret unit and the straightness of the optical axis depends highly on the correct mounting of the turret plate also the tightening sequence of the clamp mounting bolts is very important.

During tightening the clamp bolt (1) move the turret unit (turret plate) on its upper part carefully toward and away from the scanner unit until no movements can be experienced; then tighten the clamp bolt (2); only this way, the locating surface will fit correctly. The sequence of the bolts (3) and (4) is less important because these bolts are near to each other.

 

1.      Insert the (new) reflector turret unit and place the supporter tube, then insert the supporter bolt (if MIDI).

2.      Tighten the bolts in the sequence from 1 to 4 as shown.

3.      Connect the turret stepper motor cable FCJ-1.

4.      Mount the camera tube.

5.      Tighten the supporter tube bolt.

 

 After the new turret unit was mounted, all the adjustments should be checked and the appropriate parameters of the file “MicroscopeConfiguration.ini” sections [ReflectorTurret] must be updated, and the file must be saved.

 

 

 

 

 

 

 

 

 

 

 

Optical components

 

 

                         Exciting light source adapter mounting

 

The FL light source mounting interfaces and connects the appropriate adapter via a bayonet lock mechanism; with this solution, the light source may be mounted or dismounted very quickly and allows the connection of different types of light sources.

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

                         Adapters

 

Momentarily two types of adapters are available, the Lumencor® adapter and the X-Cite® type adapter.

 

 

       Lumencor SPECTRA light engine

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

                   EPI-fluorescent illumination unit

 

The aperture stop (a metal device that limits the amount of light going through the system), the luminous field stop (a metal device that limits the area of the visible field) and the condenser (a lens that serves to concentrate light from the exciting source that is in turn focused through the object) are situated on the “EPI-Fluorescent illumination unit”. The mechanical construction of the aperture stop and the luminous field stop is identical.

 

 

       Iris size adjustment tool”; “Handling the size adjustment tools”;

 “Position of the diaphragm”; Handling the position adjustment tools” and “Condenser”.

 

 

 

 

 

 

 

 

 

 

 

 

                   Position of the diaphragm

 

The diaphragm of the aperture stop and the luminous field stop are forced upward by a spring. By driving the adjustment bolts in or out a limited movement in the X- and Y-direction of the diaphragm can be performed; so the iris will be centered in relation to the beam. Very important in this construction is the force of the spring. If the force of the spring is too low (the spring is not inserted well or mechanical jamming between diaphragm and housing occurs) the diaphragm can not be adjusted or only in a very small range. If there is any problem with positioning the diaphragm, check the easy movement of the diaphragm manually, the proper position of the spring and the drive mechanics of the adjustment bolts.

Take into account, that the “Aperture size” or the “Luminous Field size” tool respectively can prevent the appropriate diaphragm from movement, even if it’s fixing is tightened!

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

                   Condenser

 

By loosening the mounting bolt on both sides of the condenser the focus position can be adjusted by moving it toward or away respectively from the luminous field stop. The condenser is mounted perpendicular to the fluorescent light beam. If the condenser’s focus is reached, a sharp view of the iris of the luminous field stop is visible on the live view mainly if the luminous field size is smaller then the image size of the camera.

  

·     Keep the lens surfaces of the condenser clean

 

 Cleaning optics” and Condenser

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

                   Mirror

 

The mirror is mounted at an angle of 45 degrees to the light beam and reflects the excitation light to the filter block.

 

             Cleaning optics”.

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

  

 

 

                   Filter block

 

The filter sets for fluorescent scan exist in various filter combinations to filter the light of a specific wavelength to excite the fluorescent stain of the tissue (Excitation filter) and to filter the relevant, emitted light of the stained tissue (Emission filter). The beam splitter reflects the shorter light wavelength during the light with the longer wavelength passes thru it. The filter sets are assembled to a filter block or filter cube. The wavelength varies in the range between ultra violet excitation (350 nm) - blue emission (450 nm) and orange excitation (600 nm) - deep red emission (690 nm).  A wide spectrum of filter sets or filter blocks is available from major microscope manufacturers via product number. If you are self assembling the filter set into a block, take care on the positions where the filters are mounted. The Emission filter shows always to the camera and the Excitation filter to the fluorescent light source. The Excitation filter, the Emission filter and the Beam splitter are combined for a special light wave length and therefore they must not be mixed with parts of another set!

 

·       Filter block”; Zeiss filter

·       Fluorescent_exciting

·       Matching Fluorescent Probes with Nikon Fluorescence Filter Blocks”; interactive

·       Introduction to Fluorescence Filters” (Semrock)

·       Filter block assembling

·       Setup filters” (to assign colors, color channels, and filter positions)

 

 

             Cleaning optics

 

 

 

 

 

 

 

When the filter block is inserted properly into the reflector turret filter wheel, the springs are fixing the filter block in its position and no further adjustments are needed.

The “hole” in the filter block (opening without filter) shows always to the objective.

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

  

 

                   FL image path cover tube

 

The fluorescent cover tube does not contain optics, it is always used in the 10th filter position of the filter wheel; it covers the image path and is inserted and mounted like a filter block.

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

                   Virtual tissue

 

To allow analyzing of parts in the tissue (e.g. nuclei, or DNS fractions), parts can be stained with special stain. A wide range of fluorescent stains (fluorophores) is available for different markers. Each stain is excited by a special wave length of the excitation light and emits light in another, relevant wavelength. One tissue can be stained with more than only one stain (fluorophore), so different parts of the tissue can be visualized in different colors at the same time.

 

To reduce the exposure time of the camera and to produce a high quality of the virtual fluorescence tissue, the used filter block must match the excitation wavelength (the source wave length to excite the stain) AND the emission wavelength (the emitted wavelength of the stain) also. Furthermore, the emitted wavelength of the exciting light source must be able to excite the stain in its wavelength.

 

To produce a high quality of the virtual fluorescent tissue and to reduce the exposure time during fluoresce scan the following parameters are very important:

          1) The characteristic of the exciting light source (emitted wave lengths)

          2) The characteristic of the used filter block (exciting and emission wave length) and

          3) The characteristic of the used stain (exciting and emitted wave length).

 

The best virtual tissue quality (and the shortest exposure time also) will be reached if all the characteristics are optimal met, otherwise the exposure time will rise up and the virtual tissue becomes more poor.

If the wave lengths of one component differ too much, the scanned quality is very poor or even bad!!

 

       Fluorescence Microscopy!

                        Fluorescent_exciting

 

 

 

             Cleaning optics

 

 

 

 

 

 

 

 

 

Adjustment tools

 

                   Autocollimator

 

An autocollimator is an optical instrument for non-contact measurement of angles. They are typically used to align components and measure deflections in optical or mechanical systems. An autocollimator works by projecting an image onto a target mirror, and measuring the deflection of the returned image against a display with a scale, either visually or by means of an electronic detector. A visual autocollimator can measure angles as small as 0.5 arc seconds.

 

Precautions         

Never look directly into the beam of the autocollimator!

The LASER beam has often a power of 0.1W only, but this is enough to harm the retina of the eye!

 

 

 

                   Dovetail ring adaptor

 

The dovetail ring adapter is used to interface the autocollimator to the adapter for X-cite type fluorescent exciting sources and the check camera.

 

 

 

 

 

                                      FL adapter for X-cite-type light sources

 

This adapter is used to connect the adjustment tools to the turret unit, because the tools are equipped with a dovetail ring adaptor.

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

                   Filter blocks

 

For adjustments, a green filter block is necessary. Nevertheless, the finished adjustments should be checked with the filter set of the user.

 

 

                   FL light source

·       The light source is used to illuminate the stained tissue during the fluorescent scan process. Depending on the light source, found by the user, the X-cite® type light engine or the Lumencor SPECTRA light engine® is used to define the luminous field size.

 

 

 

 

 

 

 

 

 

 

 

 

                   X-Cite type FL light source

 

Precautions

Never look directly into the beam of the fluorescent light source! The lamp emits also ultraviolet light with very high intensity. To prevent your eyes from harm (damage) use always sun glasses with a high filter factor of UV light if the fluorescent light source is switched on and you are adjusting the beam even if the cover of the turret unit is removed. For further precautions please, refer to the manual for the fluorescent light source you are using!

 

·       This type of light sources generates all exciting wave lengths at the same time; a white light beam is created. With this light source only single band filters can be used.

 

       Fluorescent exciting

 

 

 

 

 

 

 

 

                   Lumencor SPECTRA light engine®

 

Precautions         

Never look directly into the beam of the fluorescent light source! For further precautions please, refer to the manual for the fluorescent light source you are using!

 

·       This light source generates monochromatic exciting wave lengths; the desired wave length can be selected by software. With this light source single band or multi band filters can be used likewise.

 

       Lumencor SPECTRA light engine®

                        Fluorescent exciting

 

 

 

 

 

 

 

 

 

 

 

 

                   Check camera (optional)

 

The check camera (VRmc-8+ PRO) and the objective (TAMRON 23FM16SP), together with the auxiliary illumination it is used to make the position and the size of the aperture stop iris visible; adjustment details are visible on the screen.

 

·       The mounting with a dovetail ring adaptor allows the use of the check camera for the PMIDI and the PSCAN type scanners also.

 

Requirements

To make the aperture iris visible on the screen by the use of this camera “VRmagic”, the “VRmagic Cam Lab” is required!

Setup in the software version 1.16         VRmUsbCam DevKit for Windows (x86) 3.15b.msi

 

Setup in the software version 1.15         VRmagic USB Camera Development Kit 3.13g.msi

 

·       The camera uses an USB 2.0 port, otherwise, if the transfer rate of 480Mb/s can not be reached (USB1.1 or lower), the camera window will disappear automatically after some seconds, without any notice!!

 

 

 

 

 

 

 

 

 

 

 

Check camera mounted

 

Mount the check camera onto the X-Cite® type adapter and connect it to any USB 2.0 port. Memorize the serial number of the camera.

 

       The preview camera VRmagic” and “The program CamLab

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Adjustment techniques

The adjustment of the FL reflector turret unit includes:

 

The appropriate adjustment should be done:

 

 

 

 

If an error of the type named above occurs please, switch off the power supply and remove all the filters from the filter wheel manually. If any filter is inserted wrong one of the named behavior can occur also. Check also for unwanted / unexpected mechanical parts inside the turret unit which can inhibit the wheel from correct movement. After all filters are removed check the mechanical drive again with the Low Level Service part of the service program. Check the home positions Home1 and Home2; and check the 1st filter position, compare it with the appropriate parameter of the file “MicroscopeConfiguration.ini” section [ReflectorTurret].

 

 

 

                             Check and adjust the belt tension

·       If the movement range of the belt leading roll 1 is not enough to reach the required tension, increase the tension of the belt with the leading roll 2 and then adjust the tension with the leading roll 1.

 

Adjust the tension of the belt if:

 

1.      Loosen the belt leading roll 1 until it becomes moveable.

2.      Move the mounting bolt downward with a force of about 100 - 200gram and tighten the belt leading roll 1 mounting bolt.

3.      Rotate the filter wheel some revolutions forward and backward (with the service program).

 

The filter wheel and the motor moves correctly, if the movement sound is scarcely audible.

Check the backlash of the filter wheel also; a backlash should not be experienced.

Check also the straightness of the belt between the belt wheel and the filter wheel.

 

 

 

 

 

 

 

 

                   Check the backlash of the filter position

 

Try to move the filter wheel manually forward and backward a little bit.

 

       Check or adjust the belt tension

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

                   Loosen or tighten the belt wheel’s clamp

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

                   Position of the belt wheel’s clamp

 

§                     Rotate the stepper motor axle with the service program until the belt wheel clamp’s fixing bolt can be reached thru the hole 1 and loosen the belt wheel’s clamp.

§                     Set the stepper motor to the position Home1, fix the filter wheel by hand against rotation (so the belt wheel can also not move) and then rotate the belt wheel’s clamp to the position where the fixing bolt can be reached thru the hole 1.

§                     Check the result; repeat the procedure more times until all positions are correct.

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

                   Find the correct belt wheel’s position

 

The belt wheel position on the motor axle is defined by the position of the belt leading rolls (move the belt wheel on the rotor axle) and the Home2 state (rotate the belt wheel in relation to the rotor axle).

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

                   Adjust the Home sensor position

 

 

If the position of the home sensor was modified, check the acting range.

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

                   Find the Home position of the filter wheel

 

 

The Home position of the mechanical drive is defined by the active position of the sensor Home1 inside the motor during the permanent magnet of the filter wheel stays over the external sensor (the item “Home sensor is activated).

 

1.      Start the service program “SlideScannerService.exe” and switch active the section “reflector turret”.

2.      Rotate the filter wheel by hand or with the service program until the filter position 6 stays in the filter insert position.

3.      Move the filter wheel forward or backward with a step number of 300 steps until the external sensor shows the active state.

4.      To loosen the belt wheel clamp, may be the belt wheel has to be rotated to the position, where the belt wheel clamp’s fixing bolt can be reached by the 1.5 hex key wrench; count and memorize the done steps.

5.      Loosen the belt wheel clamp.

6.      Move the stepper motor the memorized number of steps in opposite direction; the external sensor should show the active state again.

7.      Prevent the filter wheel from movement by hand and press the button Home1, perhaps 2 or 3 times.

8.      Rotate the belt wheel’s clamp until the fixing bolt can be reached thru the drillings 1 or 2.

9.      During these actions only the belt wheel’s clamp should be rotated and the motor axle should rotate about a half revolution maximal; the filter wheel together with the belt wheel stays fixed, they should not move!

10. Rotate the belt wheel clamp into the appropriate position.

11. Release the filter wheel and go forward or backward as necessary until the clamp fixing bolt can be reached with the hex key wrench and tighten the belt wheel clamp.

12. Press Home1, 2.

 

Check the home position of the mechanical drive

 

The home position is correct, if:

·       Press the buttons Home1 and Home2 for the turret motor. The home position is found inside of 1 revolution of the filter wheel.

·       The external sensor acting range is correct.

·       The belt wheel clamp’s fixing bolt can be reached if the mechanical drive stays in Home1,2 (recommended; not required).

 

 

                       Check or adjust the external sensor acting range

 

As discussed above, the acting range of the external sensor is more 100 steps.

We assume, the Home position of the filter wheel is found; the filter wheel can be set to Home2 in one revolution maximal.

1.      Set the turret filter wheel with the service program to the position Home1,2 (B), the result is shown in the figure (2).

2.      Type in a step size of 50 steps (A).

3.      Move the filter wheel in positive direction (D) until the “Home sensor” shows the inactive state first time; figure (3).

4.      Memorize the number of steps.

5.      Set the turret filter wheel to the position Home1,2 (B).

6.      Move the filter wheel in negative direction (C) until the "Home sensor” shows the inactive state first time; figure (1).

7.      Compare the number of gone steps with the memorized number of steps.

8.      If there are more then 50 steps difference, adjust the rotation position of the belt wheel more precise.

9.      Set the turret filter wheel with the service program to the position Home1,2 (B), the result is shown in the figure (2).

10. Check the acting range again, repeat from step 1.

11. If the belt wheel position was altered, “check or find the first filter position” again.

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

                   Check or find the 1st filter position

 

 

To find the first filter position

 

A.    Mount the autocollimator with the dovetail ring adapter to the fluorescent light input connector and switch it on with the highest intensity.

 

B.    Press Home 1 and Home 2.

 

C.    After the filter is inserted, bring the filter into the light path by pressing 5 times the button 3200 forward for the turret filter unit.

 

D.    With the 100 steps button go forward or backward until the LASER beam appears on the scale.

 

E.    Select a step size of 10 steps and find the center of the scale by pressing the button without step number.

 

F.    By reducing the intensity of the laser beam the reached position can be checked better; see “Centered 2”.

 

G.   If the center is reached, subtract 16000 from the Filter step number shown for the turret unit and use the found value to update the value of the parameter “StartingMotorPosition” of the file “MicroscopeConfiguration.ini” section [ReflectorTurret] and save the file.

 

H.    Switch off the autocollimator and dismount the dove tail ring adaptor.

 

 

 

 

 

 

 

                   Procedure to adjust the exciting path

 

This adjustment is done to reach the optimal exciting for the fluorescent FOV. The “Aperture position” and the “Luminous field position” will be adjusted first to the center of the beam. If the adjustment is optimal, the brightest part of the light beam is used. With the tool “Aperture size” the diameter of the beam is limited; this eliminates unwanted reflections, otherwise in a limited range it has affect in brightness and therefore reflects to the cameras exposure time. With the tool “Luminous field size” the diameter of the visible field of view is defined. The adjustment should be done with the parts starting from the light source in direction to the tissue.

 

 

 

1.     Connect the fluorescent light source like „X-Cite® Series 120” or equivalent to the reflector turret unit of the scanner and switch it on. For user information and precautions of „X-Cite® Series 120” or equivalent light sources please, refer to the users manual of the product you are using.

 

 

 

 

2.     Loosen the fixing bolts of the positioning tools for the “Aperture” and the “Luminous field”. For the other two tool bolts the fixing bolts are situated in the opposite position, from below.

 

3.     Loosen the fixing for the “Aperture size” and the “Luminous field size”.

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

                   Aperture size and Luminous Field size tool with fixing

 

Before you start to adjust the aperture position or the luminous field position, the fixing of the “Aperture size” and “Luminous field size” tool respectively should be loosened, otherwise it blocks the diaphragm movement, because the force of the spring is not enough to move the iris. By removing the fixing, the appropriate adjustment tool can be dismounted, if necessary.

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Fluorescent light and image path

 

Precautions:  Never look directly into the beam of the fluorescent light source! The lamp emits also ultraviolet light with very high intensity. To prevent your eyes from harm (damage) use always sun glasses with a high filter factor of UV light if the fluorescent light source is switched on and you are adjusting the beam mainly if the cover of the turret unit is removed. For further precautions please, refer to the manual for the fluorescent light source you are using!

 

 

 

             Adjustment images

 

The aperture diaphragm illuminated with the fluorescent light source.

 

If the cover of the turret unit is removed, you can see the beam on the iris, if this is not fully opened.

With the “Aperture positioning tools” bring the iris into the centre of the beam. Check the position by varying the aperture size tool. The center of the beam is reached, if the cut part of the beam is illuminating the aperture iris evenly. 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

To adjust the aperture if the cover is mounted, you can use the bright field illumination.

 

Insert a sample, open the mechanical shutter, bring the filter block into its position and switch on the bright field illumination.

 

·       In the P250 please use any kind of auxiliary illumination; the flash frequency of the flash light source is too slow in the service program.

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

If you are looking now into the “Fluorescent light source connector” and the luminous field size is fully open, you can see the aperture of the objective and by moving the aperture open or close tool you see the position of the aperture iris. Adjust the iris position to the centre of the beam. By carefully varying the size of the aperture, you can check the behavior of the iris. The movement should be evenly in all directions related to the center of the beam.

 

·       Adjust the aperture size so, that the aperture of the objective is evenly not cut.

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

If the cover is removed and you are looking onto the luminous field diaphragm on the mirror side during adjustments, some times you can see that the reflected beam (reflected by the filter block) does not meet the hole of the iris. In these cases, may be the filter position is not correct (if the reflected spot is up or down in relation to the iris and no autocollimator was used; see the image on the right) and / or the position of the luminous field diaphragm is incorrect (if the reflected spot is not centered in relation to the iris).

 

Always adjust the “first filter position” before the adjustment of the diaphragm position will be done.

Check the position of the “Luminous field size diaphragm” next.

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

In ideal cases you can see somewhat like this. The field size is in the center; the filter block position is correct and the luminous field diaphragm is centered; the reflected spot illuminates the edge of the iris hole evenly.

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

                   Adjustments

 

Before you start to adjust the aperture position or the field position, the fixing of the “Aperture open or close” or “Luminous field size open or close” tool respectively should be loosened, otherwise it blocks the diaphragm movement, because the force of the spring is not enough to move the iris.

·      By removing the fixing, the appropriate adjustment tool can be dismounted, if necessary.

 

 

 

 

Check or adjust the luminous field stop position

 

The check of the luminous field stop position is done as described in the chapter “Check the filter block positions”, see later!

 

 

Prepare the FOV for the adjustment

 

1.      With the program SlideScanner.exe produce a live view with the option bright field scanning and the tab “Focus”. The tissue type can be a “normal” tissue, it is not important that the tissue is not prepared for fluorescent scanning (only the exposure time for the camera must be increased more!).

 

2.      Use a well visible FOV where the corners of the FOV have tissue also. This becomes important if we adjust the field size. Adjust the focus position and memorize it.

 

3.      Kill the program “SlideScanner.exe” with the task manager.

 

Prepare the turret unit for the optics adjustment

 

4.      Start “SlideScannerService.exe” and “Low Level Service”.

 

5.      Switch active the service part for “Reflector turret” and “Focus” and press Home1 and Home2 for “Filter” and “Focus”.

 

6.      Go to the first filter position; the value of steps is given by the parameter “StartingMotor Position” of the file “MicroscopeConfiguration.ini” section [ReflectorTurret]. In position 6 of the turret wheel insert the filter block and go forward 5 positions. If all is correct, the numerical value in the field “Filter” of the service program shows the sum of 32000 (5 times 6400 steps) + the value of the parameter “StartingMotorPosition”.

 

7.      In the “Focus” part of the service program close the shutter; use the value of “FocusDeviceMax” in the file “Microscope Configuration.ini” section [HardwareLimits].

 

8.      Open the aperture intensity and luminous field size to maximum. Start the driver program for the marlin camera “AVT SmartView,exe” and increase the shutter in the dialog “camera settings” until the FOV becomes visible. In the menu “View” select the option “resize the picture to the screen”. Now go backward with the focus stepper (do not press Home1 or Home2!) from the focus position “FocusDeviceMax” to the memorized, actual focus position. The FOV or a part of it should be seen in focus and the mechanical shutter stays closed. Switch off the bright field illumination. Adjust the shutter value for the camera and the focus position so, that the illumination and the focus of the FOV is correct.

 

Adjust the aperture position

 

9.      Prepare a sample for bright field illumination, insert the filter block and bring it in the fluorescent scan position. If the fluorescent light source is disconnected, you can see a light spot and the aperture iris in the light source connector. The right aperture position is found if the beam is in the centre of the aperture iris. The adjustment is done with the “aperture position” tools. You can check it by observing the iris in relation to the objective aperture. By carefully opening or closing the size of the aperture, you can check the behavior of the iris. The movement should be evenly in all directions related do the center of the beam. Tighten the fixing for the tool bolts.

10. Remember, that the “Aperture intensity” tool can deform the result of the position; therefore, during adjustment the intensity tool should be moved up and down from time to time, at least, before the adjustment seems to be finished.

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Adjust the luminous field position

 

11. The right “Luminous field position” is found if the beam is in the centre of the luminous field iris. The adjustment is done with the “Luminous field position” tools. You can check it by observing the iris pupil in relation to the beam. You can see the reflected light of the beam from the condenser side. If the pupil border is evenly illuminated by the beam, the adjustment is finished. You can see the tissue in the centre of the screen, if the luminous field size is nearly closed.

12. Now open it carefully until the screen border is reached by the tissue and check the luminous field position again. If the position is correct, the not illuminated parts in the corners should be equal in size. A fine adjustment of the luminous field position can be done now, if necessary.  Tighten the fixing for the tool bolts.

13. Remember, that the “Luminous field size” tool can deform the result of the field position; therefore, during adjusting the field position, the field size tool should be moved up / down from time to time, at least, before the adjustment seems to be finished.

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

                Check the filter block positions

 

·         For correct exciting of the FOV during fluorescent scan the insertion of the filters in all used positions should be checked / adjusted.

·         For this check use always the same filter cube in each position of the filter wheel.

·         During this checks, the field size diaphragm should be fully closed.

 

1.     Start the scan program “SlideScanner.exe”.

2.     Select “Microscope settings” in the menu “Options”.

3.     Connect and switch on the fluorescent light source.

4.     In the tab “Base Settings” (1) select the desired filter position; e.g. “Position 1” (2).

5.     Insert the filter cube into the filter wheel, position 1.

6.     Check “Enable position” (3).

7.     Set the radio button “Fluorescent” (4).

8.     Select the tab “Camera Rotation” (5).

9.     Load a Magazine (6) and insert a slide with (any) tissue (7) (in the first filter position only).

10. Check “Live view” (8).

11. Find a FOV inside the tissue and adjust the focus position (9) (in the first filter position only).

12. Set the “Auto Expo”sure time (10) (in the first filter position only).

13. Close the tool “Field size”; only a small circle should be seen, nearly in the middle of the red cross.

Remark: If the deviation of the filter block axis in relation to the optical axis is very much, may be you can not see the tissue (the screen is fully black). In such cases open the luminous field size until the tissue becomes visible. Adjust the filter position parts as described in “Reduce the deviation from the center” then close the tool “Field size” again. Repeat this procedure, until the center of the red cross is met inside the tissue and the tool “Field size” is fully closed.

14. Make a screenshot with “Print Screen” and save it; e.g. with “Irfan view”; the file name should be the filter position.

15. Switch off the live view, go back with the tab “Base settings” (11) and remove the filter cube.

16. Select the next filter position; e.g. “Position 2”, insert the filter and repeat from step 6 logically.

17. Execute the screenshots in all (desired) filter positions.

18. Analyze the “Print screens”.

 

 

 

 

 

 

 

 

 

 

If the center of the red cross can be found always inside the tissue, the filter cubes are inserted well and no further adjustments are needed.

The screenshots on the right show acceptable deviations from the center.

 

 

A real result of the adjustment can be found in the slide show:       Filter positions

 

The field size of the luminous field is set to the minimum. Because the tissue is visible in the center (shown with the red cross) of each filter position, the adjustment is acceptable, but the position of the field diaphragm might be adjusted a bit more precise.

 

 

 

                   Reduce the deviation from the center

 

There are 4 possibilities to reduce the elongation of the luminous field from the center of the red cross:

 

1.      Check the backlash of the filter wheel; the tightness of the belt.

 

2.      Adjust the luminous field stop position in the filter positions 2, 5 and 8, find the optimum and check always the correct insertion of the filter block in the named positions also.

3.      Check / adjust the value for the first filter position more precise.

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

4.      Increase the pressure of the springs carefully for the filter block mounting in these filter positions, where the deviation from the center is too much.

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Adjust the luminous field size

 

·         Increase the luminous field size until the entire screen is filled with tissue. Stop the adjustment and tighten the luminous field size tool. For best results the adjustment should be done two or three times.

·         Check the correct field size after all filters are inserted; the seen FOV must not be cut.

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Adjust the aperture intensity

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

                   Check the mechanical shutter closed position

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Parameters in the file “MicroscopeConfiguration.ini”

 

 

[Version]

CurrentInifileVersion=1.9;                                                actual MicroscopeConfiguration.ini file version since MScan.exe version 1.9 or higher, do not modify

[Microscope]

SerialNumber=xxxx;                                                           a four digit number defines the serial number of the P250, so the file MicroscopeConfiguration.ini is assigned to the hardware.

MicroscopeType=3DMic9;                                                 3DMic9 means Pannoramic 250

ScanCameraType=;                                                           not used

PreviewCameraType=CVrmc_m8_pPro;                       the preview camera

BarcodeReaderType=PreviewCamera

ReflectorTurretType=RT_3DH_10Pos_Belt;                 see also: P250_turret_unit

BrightfieldLightSourceType=FlashLight2010;              see also: Brightfield light source

BrightfieldLightSourceType=FlashLight2012;              see also: “ND filter unit” and “Upgrade to the software version 1.16

ObjectiveChangerType=OC_2Pos;                                 see also: Configure …

ObjectGuideXYZType=OGXYZ_FLASH3;                     see also: P250_X_Y_stage_unit

FlashUnitType=FlashUnit_Type2;                                  see also: Flash light source

NDFilterType=NDType2;                                                   see also: ND-filter unit

PreviewLightType=PreviewLightUnitType_Type2;      see also: Preview illumination unit

PowerSwitchBoardType=PowerSwitchBoard_Type1; see also: Power distribution & switch board

 

 

 

[HardwareLimits]                                                           The number of steps is always defined without mechanical jamming

 

 

ReflectorTurretMin=-300;                                                not used in the belt driven version

ReflectorTurretMax=58700;                                            not used in the belt driven version

 

[ReflectorTurret];                                                 at the end of the file

StartingMotorPosition=-30;                                             here named as “First filter position”.

MotorPositionDelta=3200;                                              Number of motor steps between two filter positions