Facility
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All Core Users: If your images were collected in our facility, please acknowledge the Confocal Microscopy Core at BWH in your paper.
Equipment Overview
The facility uses the Zeiss LSM 800 with Airyscan confocal system on a Zeiss Axio Observer Z1 Inverted Microscope. It is also equipped with an Axiocam 503 Monochrome Camera for conventional immunofluorescence. It performs:
- Conventional epifluorescence microscopy, bright field, phase contrast, and DIC/ Nomarski using monochrome camera.
- Confocal immunofluorescence, co-localization study, 3D render & reconstruction, Photobleaching studies, FRET, etc.
Live cell imaging using a built-in microscope environmental chamber with heat and CO2 control.
A motorized stage allows the capture of multiple data points during one time-lapse experiment.
Stage inserts hold 35mm glass bottom dishes as well as multi-well glass bottomed plates.
For more information about compatible fluorophores, confocal microscopy, immunofluorescence, etc. please visit the our guides, or websites listed in the links.
Core Rates
New Rates - Effective October 1, 2021
The Confocal Core Facility now services researchers in BWH only.
- For confocal, $65/hour for RIA division, $95 for other divisionss
- For widefield, $40/hour for RIA division, $55 for other divisions
Live cell imaging is charged at the same rate.
Minimum reservation time is 30 min. Time is billed in from the start time of the reservation, until the end of the actual check-out time in 30 min increments.
Cancellation Policy
Reservations may be cancelled prior to the scheduled start time. A 1-hour fee may be charged for cancellation within 48 hours for tech-assisted use, or 2 hours for independent use, of the reservation. Please contact manager if you can't cancel by yourself.
If you cancel your reservation, it is your responsibility to make sure the equipment is off if a previous user left it on for you.
Missed appointments or appointments cancelled after the start time are charged the full scheduled time.
Hours of Operation
8 a.m - 6 p.m., Monday to Friday
Reservation must be made 2 hours ahead for independent users, and 72 hours ahead for tech-assisted users.
Location
- Hale Building, Rm. 6032C
- 60 Fenwood Road
- Boston, MA 02115
Equipment Specifications - Confocal and Widefield
| Confocal | Conventional Widefield | |
|---|---|---|
| Image Acquisitions | Zeiss LSM 800 with Airyscan Confocal Laser Scanner | Axiocam 503 Monochrome Camera |
| Key Feature | Obtains optical thin sections to minimize out of focus fluorescence. More likely to photobleach than widefield. | Produces the
brightest images; suitable for thin samples. |
Software |
Zen 2.6 software with colocalization module | Zen 2.6 software |
| Light source | 4 channels: 405, 488, 561, and 640 nm laser lines | Software controlled metal-halid light source HXP 120 V |
| Color channel | 8 sequential confocal fluorescence channels, up to three parallel confocal fluorescence channels |
4 fluorescence filters sets: #10, EX: BP 450-490, EM: BP 515-565, For A488, cy2, GFP #20, EX: BP 546/12, EM: BP 575-640, For A546, cy3, Rhodamine #50, EX: BP 640/30, EM: BP 690/50, For A647, cy5, APC #49, EX: G 365, EM: BP 445/50, For DAPI, Hoechst, BFP 1 transmitted light/DIC channel |
| Resolution | 4 × 1 to 6,144 × 6,144 pixels (Airyscan max. 4,096 × 4,096 pixels) |
1936x1460 pixels |
| Speed | Up to 8 images /sec (Airyscan up to 1.6 images / sec) with 512 × 512 pixels | 38-76 frames /sec |
| Intensity control | Automatic or manual adjustment for each laser and channel | Automatic or manual exposure time, 250 μs to 60 s |
| Image bit depth | 8-bit and 16-bit | 14 Bit / Pixel |
| File Format | czi,tiff,avi etc.(Image J can also open .czi file with proper plugin) | czi,tiff,avi etc.(Image J can also open .czi file with proper plugin) |
| Pinhole | Master pinhole with preset size and position;can be adjusted as desired for multitracking and short wavelengths (such as 405 nm); automatic alignment. |
|
| Time-lapse recording | Up to 4 color channel live imaging Multiple data point acquistion using programmable stage |
One channel live imaging. |
| FRET/FRAP | Yes |
Not possible |
Core Policies
All users must read, understand, and agree to follow the policies detailed below. Thank you.
All Core Users: If your images were collected in our facility, please acknowledge the Confocal Microscopy Core at BWH in your paper.
Types of Users:
- New User: All new users must contact Dr. Jian Li for training, as described below.
- Tech-assisted: After taking training course, user operates microscope, under supervision of manager.
- Independent: Responsible for startup, shutdown and proper care of microscope during reservation. New users wanting independent status must take training course, and FIRST TIME USE CAN ONLY BE TECH-ASSISTED with no additional charge.
Training Course: One hour confocal fee will be charged for mandatory training. It generally consists of 1) a conversation with the manager to discuss the project and appropriate fluorophore choice and sample preparation and 2) a 2 hour training session on the microscope.
General policies:
- 1. Current users CANNOT provide initial training to new users. This must be done through the facility. No one is allowed to work on the microscope that is not a registered user unless arrangements have been made in advance.
- 2. All Users must Check In/Out form on the desktop computer. Please note any concerns in the comments section of the check out form, and contact Jian Li on Tues and Thurs, or Jia-Shu Yang on Mon, Wed and Fri, if you think it requires urgent attention.
- 3. Use oil sparingly on oil immersion lenses and remove oil from objectives after use by blotting the objective with lens paper. One drop is sufficient for several cover slips. Excessive use of oil and/or failure to clean up oil will cause damage to the objectives – it runs down the sides and seeps into the objective lens. This damage is costly to repair. Do NOT use Kimwipes on objectives.
- 4. Lower the objectives when you are finished. The objectives will not clear the stage if the objective wheel is turned while in the raised position – it’s an issue with many inverted microscopes. To minimize the chance of this happening we ask that every user always leaves the objectives lowered and to lower them when changing from one magnification to the next.
- 5. If anything breaks in contact with the objectives, immediately stop. Do not attempt to clean broken glass off the objective. Contact Jian Li on Tues and Thurs, or Jia-Shu Yang on Mon, Wed and Fri, and we will take care of it.
- 6. Please don’t touch any of the microscope parts with "do not change" or "do not touch" labels. If you are not sure, please contact Jian Li on Tues and Thurs, or Jia-Shu Yang on Mon, Wed and Fri, before making any adjustments.
- 7. Check the schedule to see who is on after you – and leave on or turn off the system accordingly. As a general rule, leave the system ON if there is someone signed up within 2 hours after you. If the next person is more than 2 hours away, or if you are the last person of the day you must turn the system off. Otherwise, penalty fee will be charged up to 4 hours. If you are not sure what to do, see the manager in Room 6016I. Instructions for shutdown and start up are on the computer table in the confocal room.
- 8. For live cell image, turn on CO2 tank first, then turn on CO2 and Temp Modul S before turn on computer.
- 9. Shut off the air tank for the floating table and CO2 tank for live cell imaging after use.
- 10. All data older than three months old may be deleted from the core computer. The confocal manager may delete data older than three months every month. The user is responsible for backing up their data periodically. The core is not responsible for any data lost in this way.
- 11. Penalty. It's the responsibility of last user in a day to turn off microscope. Failure to do so may leave microscope on for overnight, even entire weekend. A 2-hour penalty will be applied for each night.
THANK YOU FOR YOUR HELP KEEPING THE MICROSCOPE WORKING!
