Imaging and Cell Analysis Core
The CNR supports the Tufts Imaging Facility (TIF), and the fluorescence microscopy facilities within the Tufts Department of Neuroscience. We provide neuroscientists and other investigators with imaging services. We also provide access to a Leica cryostat for Tufts neuroscientists.
Reserve Microscope by signing up in the appropriate equipment calendar.
- For access to Stearns 207A, contact Alison Hochler
- For training and assistance with equipment use, contact Alenka Lovy.
Rob Jackson, PhD
Michele Jacob, PhD
Alenka Lovy, PhD
Please follow the links below or contact core personnel for information about services and fees.
The current fee structure for academic users of the confocal and TIRF microscopes is as follows:
- Leica SP8 - $50/hour
- Nikon A1R - $50/hour
- TIRF - $40/hour *
- Leica SPE - $25/hour **
- Keyence - $15/hour
* For longer TIRF experiments, $40/hour for the first 8 hours and then $10/hour thereafter.
** Because the Leica SP8 is heavily booked, we are offering a reduced rate of $25/hour on the SPE to encourage its use.
Please adhere to the following new sign-up rules for the SP8:
- Sign up for only 2 non-consecutive imaging sessions (4 hours each) per week during regular work hours: either 9-1pm, or 1-5pm;
- Once finished with those sessions, it is fine to sign up for more if time is available that week;
- If you need a longer imaging session (of 6 or 8 hours), you will be required to sign up very early in the morning or stay several hours into the evening (after 5pm);
- Sign-ups not adhering to these rules will be deleted.
Brightfield and Epifluorescence Microscopy
This facility is equipped with three microscopes: the Zeiss Axioplan upright, the Nikon E800 upright, and the Nikon TE300 inverted. Both Nikon microscopes are connected to cameras and equipped with Nikon NIS Element software for image acquisition. The Zeiss Axioplan upright is primarily used to confirm immunoreaction before image acquisition.
The facility has an automated stitching epifluorescence system, the Keyence, which is capable of imaging 4 channels (excitation at 405nm, 488nm, 561nm, 647nm) as well as brightfield and can scan three slides at a time. We also have three standard epifluorescent microscopes: the Zeiss Axioplan upright, the Nikon E800 upright, and the Nikon TE300 inverted. Both Nikon microscopes are connected to cameras and equipped with Nikon NIS Element software for image acquisition. The Zeiss Axioplan upright is primarily used to confirm immunoreaction before image acquisition.
|Keyence||Zeiss Axioplan||Nikon E800||Nikon TE300|
Four confocal microscopes are available, Leica SP8, Leica SP2, Leica SPE, and a Nikon A1R confocal.
The newest Leica SP8 microscope is attached to an inverted microscope, has an autofocusing feature and an automated stage capable of stitching or imaging multipoints, is equipped for live cell imaging, and has the following laser lines: 405nm, 457nm, 488nm, 514nm, 561nm, 594nm and 647nm. Our Leica SP2 and Leica SPE instruments are used in the upright configuration.
All Leica confocals are equipped with a spectral scanner system with a spectral range of detection from 400 to 850 nm. Excitation sources on the SP2 AOBS system include the argon laser generated 458nm, 475nm, 488nm, 514nm, a red diode 561nm, and a diode 633nm. The Leica SPE has all diode lasers including the 405nm, 488nm, 561nm, and 647nm.
The Nikon A1R confocal is attached to an inverted microscope, and has an automated stage capable of imaging multiple cells over time or stitching multiple image fields to obtain large images. It is also equipped with a stage top incubator and a Perfect Focus mechanism that supports live cell imaging and compensates for focus drift. There are two scanheads available, a point scanner (conventional) and a resonant scanner (30 frames/sec for 512x512 pixel field view). Simultaneous photoactivation/bleaching (using the 405nm laser) and imaging can be performed. Excitation lines include 405nm, 488/514nm, 561nm, and 647nm.
Whereas the Leica confocals are all capable of spectral detection where one has the ability to set an emission range, the emission filters on the Nikon collect the following ranges: 425-475nm (DAPI), 465-500nm (CFP), 500-550nm (GFP), 550-615nm (YFP), 570-620nm (RFP), 660-740nm (far red).
|Leica SP8||Leica SPE||Leica SP2||Nikon A1R||Nikon TIRF|
|Water Immersion Lenses||x||x|
The facility offers the use of Imaris for 3D image analysis, as well as Metamorph and NIS Elements for image quantification. Imaris is located in Arnold 309 and NIS Elements and Metamorph are installed on the Imaging Workstation in Stearns 207.
TIRF (total internal reflection flurorescence) microscopy produces very thin optical sections by creating an evanescent wave of excitation just 100nm above the coverslip without exciting molecules deeper within the specimen. This results in optical sections much thinner than those obtained by the confocal microscope, but specimens must be within the evanescent excitation wave and must be mounted in aqueous media. This inverted microscope is equipped with DIC and Perfect Focus, and does have a stage top incubator for live cell studies. There is a choice of two cameras, an EM-CCD and an ORCA-ER CCD.
The laser excitation lines available include 442nm, 488nm, 514nm, 561nm, and 639nm.
Single channel imaging filter sets are available for 488 nm, 561 nm, and 639 nm excitable fluorochromes, and almost simultaneous dual channel imaging for 488 /561 nm is possible. A quadruple filter cube (442/514/561/639nm) can be used for CFP/YFP/RFP/far red imaging.
Spinning Disk Confocal Microscopy
We are now offering access to the Perkin-Elmer spinning disk confocal (Ultraview LCI) operated by the Bunnell Laboratory and the Department of Immunology. This system is ideally suited to dynamic studies of proteins and ions in living cells.
This imaging system employs the Yokogawa CSU-10 spinning disk and an ORCA-ER CCD camera, enabling the rapid acquisition of full-field images. The Ultraview LCI system is mounted on a Zeiss 200M inverted microscope, and is equipped with 40x, 63x, and 100x oil-immersion lenses. Rapid changes in Z position can be generated using a piezoelectric motor, enabling the collection of consistent Z-stacks over time. Three lasers provide 5 excitation lines: 442nm (HeCd, 14mW), 488nm (Ar, 35mW), 514nm (Ar, 43mW), 568nm (KrAr, 14mW), and 647nm (KrAr, 14mW). Excitation sources can be rapidly switched and independently attenuated using an acoustico-optical beam splitter (AOBS). However, the laser lines that can be used concurrently are limited by the available beamsplitters:
- 442 and 514 (CFP, YFP/FITC)
- 488, 568, and 647 (GFP/FITC, DsRed/mRFP1/TRITC, and various far-red dyes)
- 442, 514, 568, and 647 (CFP, YFP/FITC, DsRed/mRFP1/TRITC, and various far-red dyes)
Much more detail is provided on the Imaging and Cell Analysis Core page for the spinning disk confocal. Here, you will find more detailed system specifications, as well as technical tips, protocols, and sample movies. Additional images are available on the Perkin-Elmer website.
The current fee structure for academic users of the spinning disk confocal is as follows:
- $60/h for use of the instrument, when assisted by the facility manager.
- $50/h for trained users, without technical assistance.
- $150 per session for training sessions.
Please contact Alenka Lovy for information about using this microscope. We are developing a training procedure for this instrument. In the future, approved users will be able to book the microscope directly.