LASER SCANNING CONFOCAL MICROSCOPY
Contact: Joe Goodhouse at goodj@.princeton.edu
Laser
scanning confocal microscopy provides our researchers the ability to optically section
whole specimens of cells and small organisms, such as developing fly and fish
embryos, or very thick sections from brain and other organ tissues that have
been tagged with fluorescent probes. This technology provides us
with the highest light microscope resolution obtainable, giving the
scientist a clearer picture of sub cellular structures, function, and cellular
or organism architecture. We can perform this on single or multiply
labeled specimens. Our laboratory has more than 17 years experience of
confocal imaging in mammalian cells, yeast cells, Drosophila and Zebra
fish embryos and ovaries, bacteria, sections of brain and other tissues, in
both fixed and live specimens. Quantitative methods such as FRAP, FLIP
AND FRET are available. The laboratory has several software packages
for image analysis, processing quantitation and 3D image reconstruction.
INSTRUMENTATION
LEICA SP5 SPECTRAL CONFOCAL - PERKINELMER RS3 SPINNING DISK
ZEISS LSM 510
LEICA SP5 CAPABILITIES
- 5 CHANNEL SPECTRAL IMAGING SYSTEM WITH EXCITATIONS AT 458, 476,
488, 514, 561, 594 AND 633.
- SPECTRAL SEPERATION OF OVERLAPPING FLUOROPHORES
- IMAGING SPPEDS FROM 10 TO 1,000 HERTZ WITH MIRROR SCANNING
- RESONANT SCANNING AT 8,000 HERTZ [25 FRAMES / SECOND AT 512 X 512
PIXEL RESOLUTION].
- FAST Z- STEPPING WITH GALVO
Z STAGE
- LIVE CELL IMAGING AT CONTROLLED TEMPERATURES EMPLOYING WARNER
HEATING UNIT
- FRAP, FRET , FLIP AND VISIBLE UNCAGING
-
PERKINELMER SPINNING
DISK CONFOCALEICA SP5 SPECTRAL SYSTEM:
This confocal
microscope was installed in August 2005. This is a 3 channel system utilizing
an Argon / Krypton laser providing excitation at 488, 568, 594 and 667 nms.
PERKINELMER CAPABILITIES
- 3 CHANNEL IMAGING SYSTEM WITH EXCITATIONS AT 488, 568 and 647.
- FAST LIVE CELL IMAGING UTILIZING CCD CAMERA AQUISITION
- IMAGE PIXEL RESOLUTION 1024 X 1344
- FAST Z- STEPPING WITH PIEZO
OBJECTIVE MOUNT
- LIVE CELL IMAGING AT CONTROLLED TEMPERATURES EMPLOYING LIVE CELL
CHAMBER
ZEISS LSM 510 CONFOCAL
SYSTEM:
The Zeiss LSM 510 Confocal system
was installed in October 1997. The system has a 3rd generation
scanhead with CFP and YFP dichroics and filter sets added. The 510 has 4 lasers producing 6 lines
[364, 458, 488, 514, 543 and 633] for fluorochrome excitation, 4 color
detectors and a transmitted light detector.
THE 510'S CAPABILITIES
- SIMULTANEOUS IMAGING 4 CHANNELS WITH COMBINED MERGED IMAGE
VIEW
- UV IMAGING INTO LIVE SPECIMENS WITH C-APO WATER IMMERSION
OBJECTIVES
- IMAGING OF CFP / GFP / YFP IN LIVE SPECIMENS FOR TIME
COURSE STUDIES AT AMBIENT AND INCUBATOR TEMPERATURES AND CO2
- YFP AND CFP IMAGING IN FIXED AND LIVE SPECIMENS
- SEQUENTIAL AND BI-DIRECTIONAL SCANNING TO AVOID CHANNEL CROSS TALK
- IMAGE RESOLUTION FROM 256 X 256 UP TO 2048 X 2048 PIXELS
- IMAGE PROCESSING IN 3D, STEREO, OR 2 1/2 D FOR
HISTOGRAM OVER AREA
- UV LASER ABLATION - FRAP STUDIES - FRET
- CAGE RELEASE OF CPDS. & FLUOROCHROMES
-
! CLICK ON IMAGE GALLERIES!