Division of Biology and Medicine
BioMed Core Facilities
Zeiss Axiovert 200M Fluorescence Microscope (LMM)
Imaging of samples labeled with fluorescent indicators
Zeiss Axiovert 200M Fluorescence Microscope (LMM)
Imaging of samples labeled with fluorescent indicators
Overview
Laboratories for Molecular Medicine, Room 111
The Zeiss Axiovert 200M Fluorescence Microscope is a motorized inverted microscope with a Zeiss Mrc5 color camera and a Roper CoolSnap HQ monochrome camera controlled Axiovision 4.8.2 software. This microscope is well equipped for transmitted light microscopy and fluorescence microscopy.
Operation
Objectives:
|
Mag |
Correction |
NA |
Phase |
DIC |
WD (mm) |
Coverslip # |
|---|---|---|---|---|---|---|
|
5x |
Plan-NEOFLUAR |
0.15 |
Ph1 |
- |
13.6 |
0.17mm #1 1/2 |
|
10x |
Plan-NEOFLUAR |
0.3 |
Ph1 |
DIC II |
5.5 |
0.17mm #1 1/2 |
|
20x |
Plan-NEOFLUAR |
0.5 |
Ph2 |
DIC II |
2.0 |
0.17mm #1 1/2 |
|
40x |
Plan-NEOFLUAR |
0.75 |
Ph2 |
DIC II |
0.5 |
0.17mm #1 1/2 |
|
40x Oil |
Plan-NEOFLUAR |
1.3 |
Ph3 |
DIC III |
0.2 |
0.17mm #1 1/2 |
|
100x Oil |
Plan-NEOFLUAR |
1.3 |
Ph3 |
DIC III |
0.2 |
0.17mm #1 1/2 |
Theoretical XY Objective Resolution: based on: Dxy=0.61*wavelength/NA
|
Mag |
NA |
DAPI |
FITC |
RHO |
CY5 |
|---|---|---|---|---|---|
|
5x |
0.15 |
1870 nm |
2175 nm |
2520 nm |
2930 nm |
|
10x |
0.3 |
935 nm |
1090 nm |
1260 nm |
1465 nm |
|
20x |
0.5 |
560 nm |
650 nm |
755 nm |
880 nm |
|
40x |
0.75 |
375 nm |
435 nm |
505 nm |
585 nm |
|
40x Oil |
1.3 |
215 nm |
250 nm |
290 nm |
340 nm |
|
100x Oil |
1.3 |
215 nm |
250 nm |
290 nm |
340 nm |
Transmitted light microscopy: The Axiovert 200M has DIC and Phase Contrast on all objectives. The high resolution color camera can be used for imaging fixed samples (e.g. histological preparations) and for time-lapse recordings of living cells. A stage heater is available for imaging living cells at a set temperature.
Fluorescence microscopy: The filter sets allow imaging of a wide variety of fluorescent indicators such as Alexa-Fluors, Calcium-Green, Cy2, Cy3, Cy5, DAPI, Fluorescein (FITC), FM1-43, Fluo-3, Fluo-4, GFP, CFP, YFP, Hoechst 33258, Rhodamine, Texas Red, TRITC, and others. The stage heater and the sensitive monochrome camera make this microscope particularly well suited for imaging dynamic patterns of fluorescence in living cells.
Filter sets on the Axiovert 200M:
1) DIC: Analyzer for DIC
2) DAPI: Chroma set 31000V2, Excitation 325-375 nm, Emission 435-485 nm (bandpass filter).
3) FITC: Chroma set 41001, Excitation 460-500 nm, Emission 510-560 nm (bandpass filter).
4) RHOD: Chroma set 41002c, Excitation 530-560 nm, Emission 590-650 nm (bandpass filter).
5) Cy5: Chroma set 41024, Excitation 590-650 nm, Emission 665 nm and above (long pass filter).
Also available upon request:
6) GFP: Chroma set 41017, Excitation 450-490 nm, Emission 500-550 nm (bandpass filter).
7) CFP: Chroma set 31044v2, Excitation 426-446 nm, Emission 460-500 nm (bandpass filter).
8) YFP: Chroma set 41028, Excitation 490-510 nm, Emission 520-550 nm (bandpass filter).
9) CFP/YFP FRET: Chroma set 31052, Excitation 426-446 nm, Emission 520-550 nm.
Reservations
1. Go to our facility's iLab site: ILABS
https://brown.ilab.agilent.com/service_center/show_external/5278/bioimaging_facility
2. In the upper-right-hand corner of the screen click ‘Sign In,’ and select to sign in using Brown University credentials
3. You will be directed to an authentication page where you will enter your Brown University credentials
4. Once you have entered your credentials, click the ‘Login’ button
5. You will be directed to an iLab Registration page where you will select your PI/Lab and verify your contact information.
6. Once your registration has been submitted, your PI will receive a notification that you have requested membership to their lab in iLab. They will need to approve your membership.
7. Your training status needs to be updated. This is done on a rolling basis by Eric Olson (Eric_Olson@brown.edu) and Sandra Van Wagoner (sandra_van_wagoner@brown.edu). Alternatively, you can email Geoffrey_Williams@brown.edu or Robbert_Creton@brown.edu asking for an update of the training status in iLab, listing the microscopes that you have been trained on in the Bioimaging Facility.
When registration is completed (including step 6-7), you can schedule equipment using the iLab calendars.
1. Navigate to the core page: ILABS BIOIMAGING
https://brown.ilab.agilent.com/service_center/show_external/5278/bioimaging_facility
2. In the upper-right-hand corner of the screen click ‘Sign In,’ and select to sign in using Brown University credentials
3. You will be directed to an authentication page where you will enter your Brown University credentials
4. Once you have entered your credentials, click the ‘Login’ button
5. Select the Schedule Equipment tab and click on the ‘View Schedule’ button next to the instrument of interest. Click and drag on the time frame you would like to schedule your reservation for.
6. A window will pop up that will allow you to verify your reservations details and provide payment information before saving the reservation.
When it is time to use the microscope
1. Open iLab and select 'Go to Kiosk'
2. Start end End microscope session (Start is needed to run the microscope)
Additional help
More detailed instructions can be found on the Help Site by clicking on the “HELP” link in the upper right hand corner. For any questions not addressed in the Help site, click on the “HELP” link in the upper right hand corner and submit a ticket, or email ilab-support@agilent.com.