Simplified Instructions: Zeiss Brightfield Microscope S1000
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1 Contents General Microscope Set-Up Adjust Illumination Focus Condenser Open Software Image Capture Settings Shading & Color Corrections Image Capture & Viewing Scaling and Measurements Synopsis of Other Functions Pages Simplified Instructions: Zeiss Brightfield Microscope S1000 This is an Overview Only intended to refresh specific items learned in the Hands-On Training Use only in conjunction with your own much more detailed notes taken during your training
2 1, Reserve Time on Scheduler 2, Remove Eye-Piece Covers
3 3, Power-Up Computer is Usually On (if not, turn on)
4 3, Power-Up Turn on Lamp (on Left of Stand)
5 3, Power-Up Adjust Ocular/Camera Light Path To 50:50 (middle position)
6 3, Power-Up Manually Move Desired Objective Into Position
7 3, Power-Up Move Condenser Magnifier INTO LIGHT PATH If Using 10x or Higher Objective
8 3, Power-Up Move Condenser Magnifier OUT OF LIGHT PATH If Using 5x or Lower Objective
9 4, Adjust Illumination Adjust Light Filters (recommend none, e.g. II, H) (i.e. anything where light is brightest)
10 4, Adjust Illumination Adjust Lamp Power (on Right of Stand) Recommend 3200K
11 4, Adjust Illumination Set to Frosted or Clear Glass (Frosted Recommended, which is as shown below)
12 4, Adjust Illumination Focus on your slide
13 Note: Altering Brightness Through Neutral Density Adjustment is Preferred Over Changing Lamp Power 4, Adjust Illumination Adjust Neutral Density Filters To Alter Brightness to Good Level While Viewing Sample via Oculars
14 RECOMMENDATION Each User Should Define A Standard Frosted Glass/ Neutral Density/Condenser Setting. Make note of that setting and use each time for consistent imaging. 4, Adjust Illumination Can also Adjust Illumination Level by Changing Light Path Condenser
15 5, Focus Condenser (must do for Every Objective) A, Focus Sample, use Eyepieces B, Close Aperture F (right of stand) to Create Ring On Image (see pic)
16 5, Focus Condenser (must do for Every Objective) C, Move Condenser Knob Until Margins of Ring are Sharp (see pic)
17 5, Focus Condenser (must do for Every Objective) D, Center Ring in Middle of Field Using Adjusting Screws
18 IMPORTANT You have focused the illumination light, which is different for each objective. If you change the objective, you must re-focus the condenser for the new objective. 5, Focus Condenser (must do for Every Objective) E, Once Centered and Sharp, Expand Ring (with Aperture, p 15) to Slightly Bigger Than Viewing Field
19 6, Open Software Use Windows Log-In/Password provided to you at training Click on AxioVision Rel. 4.6 icon
20 6, OpenSoftware Workarea icon open the window on the left. Each laboratory may have modified the workarea. Note: right-click on upper area permits you to set toolbars available.
21 7, Image Capture A, Click Live to view sample on screen.
22 7, Image Capture B, Use Magnification drop-down to tell software which objective is used.
23 7, Image Capture C, Select speed of Live Image update to Slow D, Focus Image on screen
24 RECOMMENDATION Do not use any Display other than Linear. Under NO circumstances do you use G Best Fit and Min/Max Settings are more for fluorescence microscopy and are not recommended here. 7, Image Capture E, Make sure Linear Image Display is highlighted.
25 7, Image Capture F, Click on Properties to open the main capture settings menu. G, Make NO adjustments under Display tab. Using Linear (already set) and next pages adjustments ensures consistency.
26 7, Image Capture H, Under Adjust tab, Click Measure to have the computer adjust exposure time for you. You also can manually adjust by typing in the desired number of msecs to be exposed.
27 7, Image Capture I, Shading Correction Open General Tab Move Slide to a field of no sample and no debris Moving to slightly out of focus can minimize presence of any debris
28 7, Image Capture I, Shading Correction Click Shading Correction Note: the slight variations in darkness (see last slide) are now even across the image.
29 7, Image Capture J, Set Color Balance Return to partial sample in field Return to Adjust Tab Focus Sample Click Interactive (changes to picking )
30 Note, after adjustment display shows equal blue, green and red. (compare to prior page) 7, Image Capture J, Set Color Balance Move mouse to empty portion of field and click Note: background color now has similar blue, green and red components.
31 7, Image Capture K, Capture Image Move to desired image field. Click Snap to capture image
32 8, Save Image Under File
33 8, Save Image Save Images only to a mapped server or to a temporary storage device. If you need assistance with server mapping, contact your IT support. Under NO circumstance do you save to the microscope computer.
34 9, Viewing Image Image captured is larger than Display. Use sliders to display area of image you are interested in. Note: Despite display, whole image will be saved, not just the visible part.
35 9, Viewing Image Can use Maximize to fill image window (see next page)
36 RECOMMENDATION Follow all the above steps prior to image collection and change nothing post-capture. If you make changes here, this image will be displayed different from your others. 9, Viewing Image Can open Properties pop-up, which Allows changes similar to those Described under live properties (p. 24)
37 10, Add Scale Bar Under Annotation, select Scale Bar
38 10, Add Scale Bar Scale Bar Appears Click on Scale Bar to move it. Note: Important to select correct magnification at collection (slide 22)
39 10, Add Scale Bar If incorrect magnification selected: A, Open Scalings under Measure
40 10, Add Scale Bar If incorrect magnification selected: B, Highlight correct magnification C, Click Apply selection to image NEVER touch anything else here as you may incorrectly alter the scalings. RECOMMENDATION Ideally you will always set the Magnification during set-up. It is unlikely you will remember this settings weeks later. i.e. use for your record-keeping.
41 11, Measurements Under Measure, select type of measurement to be conducted
42 11, Measurements Create Region of Interest (ROI) by holding down left-click of mouse Right-click mouse to close the ROI after defining the circle. Create as many ROIs as you would like
43 11, Measurements After creating all ROIs, click on Create Data Table
44 11, Measurements Apply Data to Table
45 11, Measurements Go to File Save As to store data table.
46 11, Measurements Save your data to the mapped server or your external storage device. Recommend to save as a.csv file (can be opened in Excel)
47 11, Measurements Use Window to toggle between open images or tables, or Simply click the desired Tab
48 12, Synopsis of Other Functions in Software Most Important Tab: Help Tab This is your Go-To Tab for all of your further questions!
49 12, Synopsis of Other Functions in Software Highlight each Tab to view functions not discussed in this handout. 1. File Tab
50 12, Synopsis of Other Functions in Software Highlight each Tab to view functions not discussed in this handout. 2. Edit Tab
51 12, Synopsis of Other Functions in Software Highlight each Tab to view functions not discussed in this handout. 3. View Tab
52 12, Synopsis of Other Functions in Software Highlight each Tab to view functions not discussed in this handout. 4. Acquisition Tab
53 12, Synopsis of Other Functions in Software Highlight each Tab to view functions not discussed in this handout. 5. Functions Tab
54 12, Synopsis of Other Functions in Software Highlight each Tab to view functions not discussed in this handout. 6. Annotation Tab
55 12, Synopsis of Other Functions in Software Highlight each Tab to view functions not discussed in this handout. 7. Measure Tab
56 12, Synopsis of Other Functions in Software Highlight each Tab to view functions not discussed in this handout. 8. Tools Tab Options dialog can be helpful to the User
57 12, Synopsis of Other Functions in Software Highlight each Tab to view functions not discussed in this handout. 8. Tools Tab A, Options: General
58 12, Synopsis of Other Functions in Software Highlight each Tab to view functions not discussed in this handout. 8. Tools Tab B, Options: Layout
59 12, Synopsis of Other Functions in Software Highlight each Tab to view functions not discussed in this handout. 8. Tools Tab C, Options: Folder
60 12, Synopsis of Other Functions in Software Highlight each Tab to view functions not discussed in this handout. 8. Tools Tab D, Options: Storage
61 12, Synopsis of Other Functions in Software Highlight each Tab to view functions not discussed in this handout. 8. Tools Tab E, Options: Acquisition
62 12, Synopsis of Other Functions in Software Highlight each Tab to view functions not discussed in this handout. 8. Tools Tab F, Options: Naming
63 12, Synopsis of Other Functions in Software Highlight each Tab to view functions not discussed in this handout. 8. Tools Tab G, Options: Display
64 12, Synopsis of Other Functions in Software Highlight each Tab to view functions not discussed in this handout. 8. Tools Tab H, Options: ID
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