Reflector turret unit; S_M

Gear driven

For technicians and partly for sales managers!

 

 

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

 

Contents

General

Functional overview

Construction of the fluorescent exciting and image path

Construction of the RTU

Mechanical components and construction

·         Stepper motor

·         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

 

Adjustments and checks

·         Tools, used for the adjustments

·         Find the hardware limits

·         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

 

 

 

 

General

 

The reflector turret unit is a component added to the Pannoramic SCAN and Pannoramic MIDI scanners respectively to give the possibility for fluorescent exciting and scanning of tissues. This component is not used for bright field scanning. For fluorescent scanning of tissues, light color filters are used in much variation. The filter wheel in this turret unit has ten positions, so it can contain up to 9 light filters for fluorescent scanning. If bright field or fluorescent scanning is used, in one filter position the turret tube must be inserted (default position =10). When the filters are inserted, the user can select each filter (position) by software.

 

The instructions are shown for Pannoramic SCAN; differences to Pannoramic MIDI are explained at the actual step. A significant difference between Pannoramic SCAN and Pannoramic MIDI is, that the reflector turret unit in the Pannoramic SCAN is mounted vertically during in the Pannoramic MIDI it is mounted horizontally. This difference has no aspect in functionality; the reflector turret units are identical.

 

The exchange of the entire turret unit is possible

 

·      If the stepper motor or its electronics is faulty.

·      If the shape of any part is deformed or a part is broken.

·      If the turret unit has any fault and you are unable to fix it.

 

 

Requirements

  • Service program for Pannoramic scanners (SlideScanner Service.exe)   with actual license file
  • Pannoramic SCAN and Pannoramic Viewer software (SlideScanner.exe, SlideViewer.exe) with actual license file
  • 1.5, 2.5, 3 and 5 mm hex wrenches,
  • Hardware and construction knowledge of Pannoramic SCAN and MIDI respectively
  • Deeper knowledge of handling the Pannoramic SCAN and Pannoramic VIEWER software

 

 

Attention:           Do not mix the versions of SlideScanner.exe and SlideScannerService.exe! Always use these programs with the same version number. Otherwise the SlideScannerService.exe program could produce unwanted results and SlideScanner.exe does not work correctly or even freeze!

 

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

 

       "Stepper motor"

 

 

 

 

Functional overview

 

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 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”.

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 of 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 and the tube lens 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 wave length; 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

                        Influence of the camera adapter” and “Useable resolutions of scan (main) cameras

 

 

 

Physical solution of the exciting path

  

Traditionally, the fluorescent light beam may contain all the wavelengths from ultra violet (from about 320nm) thru the visible light (about 400nm to 720nm) until the infra red spectrum (over 720nm 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 by the exciting filter. 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 illuminated (excited).

 

Important

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

 

 

In newer exciting light sources like the Lumencor® Spectra, the exciting light will be generated by power Illumination modules which produce the exciting light wavelength directly. By switching the Exciting 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.

 

       X-Cite® Series 120Q”, “Xcite®120PC_UserGuide”, “Filter block”, “Lumencor® Spectra”, “Determining the Filter Type” (Semrock)

 

                        Fluorescent_exciting

 

 

 

 

Construction of the RTU

 

1          Fluorescent light input connector

2          Aperture stop diaphragm

3          Field stop diaphragm

4          Condenser

5          Mirror

6          Filter block

7          Filter wheel

8          FL image path cover tube

9          Transmission gear

10       RTU motor

11       Field size adjustment

12       Aperture size adjustment

13       Flexible shaft for the position adjustment bolts

 

 

 

 

 

 

 

 

Components of the fluorescent RTU

 

                   Stepper motor

 

 To rotate the filter wheel of the turret unit a 2-phase stepper motor with a resolution of 3200µ-steps/revolution is used. This way, any inserted filter (or filter position) can be selected via software commands at the appropriate moment.

 

  • The motor is driven in micro steps; the revolution of the rotor is divided into 3200µ-steps.
  • The stepper motor of the gear driven reflector turret unit gets its commands via the bus cable STF-4.
  • The address of the motor electronics is 06.
  • The stepper motor is mounted to the turret plate via the “Mounting nuts”.

 

       Stepper motor

 

 

 

 

                   Gear drive

 

The gearwheel on the motor axle drives the filter wheel via the transmission gear. The mechanical drive solution allows a filter position change with exact 6400 motor steps. In other words, the motor axle does exact two revolutions 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 positions forward, i.e. the motor axle does 18 revolutions to reach the last filter position.

 

·       By loosening the transmission gear mounting and moving it away from the motor axle, the filter wheel disconnects also from the transmission gear and so the position of the limiter in relation to the transmission gear can be modified; the required hardware limit position can be found.

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

                   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 the filter block positions.

 

  • Keep the position surfaces of the filter block and the filter wheel clean and dry.
  • During insertion or removal of the filter block take care on the filter fixing springs; do the action carefully, without deforming the springs.

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

                   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 (the position “Press downward”) until the springs are even disconnected, then move the entire filter block sideward.

 

  • During the removal of the filter block, the springs on the top must not be deformed!

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

To avoid miscounting of steps a mechanical limiter is mounted on the filter wheel (the filter wheel would be able to rotate endless, but the sensor home 2 finds the home position of the mechanical construction only inside a limited number of rotor revolutions). This way, the mechanical construction has a start point (negative mechanical limit) and an end point (positive mechanical limit). The limiter is mounted between two filter positions so, that both filter positions can be reached; these are the mechanical first and the mechanical last filter position (not the 1st and the 10th filter position). If the limiter is mounted in wrong position (if it is mounted on the neighbor bolt), the limits can not be adjusted correctly! The negative limit is not, and must not be identical with the home position! The size of the limiter and its mounting position guarantees, that no motor steps are lost, if the adjustment of the mechanical drive was done correctly. The distance between the first filter position and the 10th filter position is always exactly 57600 motor steps!

 

 

 

·       The size of the limiter is designed to fulfill the Home 1 limit of the stepper motor.

 

       How to define hardware limits”, “Stepper motor implementation”, and “Construction of Home1 and Home2

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

                   Drive and limits

 

Because a limiter is mounted on the drive part of the filter wheel, the mechanical movement has a negative and a positive limit.

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

·       The accuracy of the hardware limits is 100steps

 

The number of steps from the home position to the first filter position (about +300 steps) can differ by a few hundred steps.

 

       How to define hardware limits”,

 

 

 

 

 

                         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

  • Loosen the “Aperture size” and / or the “Luminous field size” tool knob fixing.
  • Loosen the appropriate position tool knob fixing bolt(s).
  • Rotate the tool knob(s) until the desired position is reached.
  • Tighten the tool knob fixing bolt(s) to fix the adjusted position.

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

                   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 and 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 and 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

  • Loosen the tool knob fixing.
  • Move the tool knob up or down until the desired size of the aperture or the luminous field respectively is found.
  • Tighten the tool knob fixing.

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

                   Check mechanical shutter closed position

 

  • During a fluorescent FOV is visible on the screen (as live view) switch on and off the bright field illumination several times with the service program. If the shutter is in proper position the brightness of the view must not change.

 

 

       Focus unit’, and  Shutter mechanics

 

 

 

 

 

 

 

 

 

 

 

 

 

 

                   Remove the RTU (SCAN)

1.      Remove the camera tube (if exchange).

 

2.      Remove the cable STF-4 (if SCAN) from the turret stepper motor.

 

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

 

4.      Pull the entire fluorescent reflector turret unit in an angle of approximately 30 degrees to your self and upward.

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

                   Insert and mount the RTU (SCAN)

 

1.      Insert the new reflector turret unit.

 

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

 

3.      Connect the turret stepper motor cable STF-4.

 

4.      Insert the camera tube.

 

After the turret unit was changed, the adjustments described above should be checked and the appropriate parameters of the file “MicroscopeConfiguration.ini” sections [ReflectorTurret] and [Hardware Limits] must be updated, and the file must be saved. In all cases the chromatic aberration and the camera angle must be adjusted; see also “Stitching”.

 

 

 

 

 

 

 

 

 

 

                   Remove the RTU (MIDI)

1.      Remove the camera tube (if exchange).

 

2.      Remove the cable for the turret motor STG-4 (not shown).

 

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

 

4.      Remove the supporter mounting bolt; hold the turret unit, remove the supporter tube carefully and then pull the entire unit as shown.

 

Attention!     The turret unit is heavy!

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

                   Insert and mount RTU (MIDI)

 

1.      Mount the turret unit in reverse order as it was dismounted.

 

2.      Insert the turret unit, underpin it with the supporter tube and mount the supporter mounting bolt. The turret unit should be fully inserted (without a gap to the main truss)!

 

3.      Mount the clamp to the main truss and then tighten the bolts in the shown sequence.

 

4.      Connect the cable STG-4 to the turret stepper motor.

 

5.      Finally insert the camera tube.

 

After the turret unit was changed, the adjustments described above should be checked and the appropriate parameters of the file MicroscopeConfiguration.ini sections [ReflectorTurret] and [Hardware Limits] must be updated, and the file must be saved. In all cases the chromatic aberration and the camera angle must be adjusted; see also “Stitching”.

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Components and construction of fluorescent optics

 

 

                   Light source connector

 

The flexible light guide of the excitation light source is connected by the help of the collimating adaptor to the light source connector.

 

·       To reach an evenly exciting of the field of view, the flexible light guide should be guided straight about 30cm before it arrives to the collimating adapter.

 

 

 

 

 

 

 

 

 

 

 

 

 

 

·       To avoid loss of brightness, the light guide must be fully inserted into the collimating adaptor and, the light guide must be fully inserted on the lamp side also.

·       Never bend the flexible light guide excessive!

 

       X-cite120PC_UserGuide”, chapter 7 and chapter 8

 

 

 

 

 

 

 

 

 

 

 

 

                   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.

 

 

 

 

 

 

 

 

 

 

 

 

                   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 fixing bolts 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 even 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.

  • The mirror does not need adjustments.
  • Keep the mirror surface clean; see also: “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 ör 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

·       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)

 

  •  Keep the surfaces of the excitation filter and the emission filter clean; see also: “Cleaning optics

 

 

       Optical path and Field Of View

                        Fluorescent_exciting

 

 

 

 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.

 

  • The proper position of the filter block is reached, if the springs on the top are pressing the filter block onto the position surface and the springs from beside are pressing the filter block toward the gear wheel!

 

 

 

 

 

 

 

 

 

 

 

 

 

  

 

                   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 or 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!!

 

More information about     Fluorescence Microscopy!

                                               Fluorescent_exciting

 

 

  • Keep the surface of the cover slip and the surface of the slide bottom clean.

 

        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.

 

 Attention: Do not forget to remove the fluorescent shutter from the fluorescent light path; otherwise the filter wheel position can not be adjusted.

 

 Autocollimator   Wikipedia

 

Dovetail ring adaptor

 

The dovetail ring adaptor is used to interface the autocollimator and the check camera to the light source 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.

 

       X-Cite Series 120Q” and “Xcite120PC_UserGuide

                        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 280Mb/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.

 

       Preview camera VRmagic” and “Program CamLab

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Adjust filter wheel and mechanical drive

 

The relevant adjustment is done by positioning the connection of the transmission gear wheel in relation to the limiter of the filter wheel, while the stepper motor is in Home1,2 position.

 

This adjustment should be done:

 

  • If the home positions Home1 and Home2 (with the service program, Low Level Service) cannot be reached.
  • If motor steps are lost during Home1 and Home2 will be reached.
  • If moving to the last filter position causes lost motor steps (check the backlash).  
  • If moving to any filter position causes lost motor steps (check the backlash).  
  • If the gear bolt was loosened
  • If the stepper motor had been exchanged

 

If an error of this type named above occurs please, switch off the power supply and remove all the filters from the filter wheel. If any filter is inserted wrong (upside down or not correctly) this behavior can occur also. Check also for unwanted or unexpected mechanical parts inside the turret unit which can inhibit the wheel from 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; check the negative turret limit, the backlash in each filter position and the positive turret limit, and compare them with the appropriate parameters of the file “MicroscopeConfiguration.ini” sections [HardwareLimits] and [ReflectorTurret].

 

 

 

 

                   Define hardware limits

 

1.      Loosen the gear bolt and move it downward, so that there is no connection between motor axle and filter wheel, the gear is disconnected. Check the easy and soundless movement of the filter wheel manually!

 

2.      Start the program SlideScannerService.exe and Low Level Service.

 

3.      Switch active the section “Reflector turret” and press Home1 and Home2 for “Filter”.

 

4.      If motor movement stops and the home positions Home1 and Home2 are not reached, press Home1 and Home2 for “Filter” again until the home positions are reached (it can take a minute, depending from the actual position).

 

5.      Bring the filter position 6 to the top, insert a small piece of paper or aluminum foil between the cogs of the gear wheel and the filter wheel, connect the gear (move the gear bolt upward) and tighten the gear bolt so that there are about 3 cogs distance exists between the gear wheel and the limiter. The small piece of paper between the cogs of the gear wheel and the filter wheel helps to reach a small backlash (about 0.1 mm) because the Filter wheel is never exactly round or centered. Do not forget to remove the piece of paper or foil if the adjustment is finished!

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Adjust the negative turret limit

6.      Press Home1 and Home2 again.

 

7.      With the “-100 steps” button go to the negative limit until the filter wheel movement stops, jamming occurred or -1500 steps are reached. If the position of -1500 steps is reached press Home1 and Home2, loosen the gear bolt, repeat step 5 meanwhile decreasing the number of cogs in distance to the limiter by 2. If the reached step value without jamming (subtract in mind 100 steps of the shown value if jamming occurred) is between 300 and 600 steps the adjustment is O.K. If the step value is less then 300 steps, increase the distance to the limiter by 1 cog. If the step value is more then 600 steps, decrease the distance to the limiter by 1 cog. Repeat steps number 6 and 7 until the negative limit is found correctly.

 

8.      The correct negative limit is found if: -600 steps negative limiter -300 steps, without mechanical jamming. Update the parameter value of the parameter “ReflectorTurretMin” with the found value in the file “MicroscopeConfiguration.ini” section [HardwareLimits].

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Backlash of the filter positions

 

9.      Go forward from filter position to filter position 9 times and check the backlash manually in each filter position. A very little backlash should exist (less then a half cog in distance) in each filter position. Listen also to the wheels movement sound. If there can be recognized a sound of mechanical jamming or in a filter position is no backlash, increase the backlash by loosening the gear bolt, moving it carefully downward a little bit and tighten it again. Repeat this step until the size of the backlash is acceptable.

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Find the step number for the first filter position

10. The best way, to find the first filter position is the use of an autocollimator. If you don’t have an autocollimator, you can adjust the first filter position as described in the following step, the results are acceptable also. For the adjustment procedure with the use of an autocollimator please refer to the chapter “Autocollimator” after this adjustment procedure and step 9 of this procedure can be left out.

 

Press Home1 and Home2.

 

 The position, where a filter can be inserted is now the position 6, the position one is exactly in opposite position, in the light path.

a.      Possibility:

In this position insert the turret tube and place a spirit level on it. With the service program go forward by +100 steps until the level is reached. For fine adjustment you can use a step size of 50 steps. Use the found value to update the value of the parameter “StartingMotor Position” of the file “Microscope Configuration.ini” section [ReflectorTurret] and save the file. An exact fine adjustment of the first filter position is done later, if the fluorescent light path will be adjusted.

 

·       Take into account, that the spirit level has a tolerance of almost 20%; so the result will not be exact. If possible, please adjust the first filter position always with an autocollimator.

 

       Autocollimator”.

 

 

 

 

 

 

 

 

 

 

 

 

b.      Possibility, if you have no spirit level or the adjustment is done for Pannoramic MIDI:

It is recommended to do this adjustment before optical adjustments are done, otherwise the camera rotation angle must be adjusted again.

Dismount the camera tube and look inside the hole. With the service program go forward by +100 steps from Home1 and Home2 until the hole of the turret filter wheel is in the center of the mounting of the camera tube. For fine adjustment you can use a step rate of 50 steps.  Use the found value to update the value of the parameter “StartingMotorPosition” of the file “Microscope Configuration.ini” section [ReflectorTurret] and save the file. An exact fine adjustment of the first filter position is done later, if the fluorescent light path will be adjusted.

 

 

·       Take into account, that the used method is not precise; so the result will not be exact. If possible, please adjust the first filter position always with an autocollimator; see also:          Autocollimator”.

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

To adjust the positive limit

 

11. Starting from the last filter position check the number of steps, until the positive limit is reached. There should be at least +300 steps until jamming, but not more then 59100 steps absolute in relation to Home1 and Home2, otherwise do the entire adjustment again from step one. Check also the position of the limiter. If it is mounted in wrong position, the limits can not be reached correctly. The correct positive limit is found if: Last filter position +300 steps positive limiter ≤ +59100 steps absolute, in relation to Hom1, Home2; without mechanical jamming. Update the value of the parameter “ReflectorTurretMax” with the found value in the file “Microscope Configuration.ini” section [HardwareLimits] and save the file.

 

Finally

 

12. Test the adjustment. Calculate and type in the number of motor steps to test the reflector turret unit from Home1,2 to the last filter position; the number of steps is found as follows: Number of steps for the first filter position + 57600 (9x6400). Move the filter wheel from Home1,2 to the last filter position and this number of motor steps return with the unnumbered blue arrow buttons for reflector turret “Filter” to Home1,2, but do not press Home1,2. 

Execute the test forward and backward with the defined number of steps, then press Home1. If the adjustment is correct, there should not be more then 1 or 2 steps difference to Home1,2. Now press Home2 and do this test three times. If there are more then 2 steps lost, check the backlash and the adjustment again. Do not forget to remove the piece of paper or foil.

 

 

                   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.

 

Attention: Do not forget to remove the fluorescent shutter from the fluorescent light path; otherwise the filter wheel position can not be adjusted.

 

 

 

 

 

 

 

 

 

                   Check or find the 1st filter position

 

  • This adjustment assumes that the backlash of the gear wheel is adjusted correctly.
  • The Home1,2 position of the filter wheel is correct.

 

 

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 6400 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 32000 from the Filter step number (because we moved the filter wheel 5x6400steps forward) 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.

 

 

 

 

 

 

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 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!

 

 

 

                      Adjustment images

 

                      FL Aperture

 

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 aperture 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 even in all directions related do the center of the beam.

 

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

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

                      Luminous field

 

  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 on the left or on the right 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, checks

 

Check or adjust the aperture diaphragm position

Loosen the fixing bolt of the aperture size.

 

Before you start to adjust the aperture position or the field position, the fixing of the “Aperture size” or “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. 

 

 

 

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

 

 

 

Prepare the FOV for the adjustment

 

4.     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!).

 

5.     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.

 

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

 

Prepare the turret unit for the optics adjustment

 

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

 

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

 

9.     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”.

 

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

 

11.  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

14.  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.

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

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Adjust the luminous field position

15.  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.

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.

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 or 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 or 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 “To 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 also 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 and 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

 

  • Close fully the aperture intensity tool then open it carefully until the brightness is not increased more. Stop the adjustment and tighten the tool fixing. For best results the adjustment should be done two or three times.

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

                   Check the shutter closed position

 

  • While a fluorescent FOV is visible on the screen (as live view) switch on and off the bright field illumination several times with the service program. If the shutter is in proper position the brightness of the view must not change.