Equipment
Knapp
Center Microscopes
Leica TCS SP2 AOBS Laser Scanning Confocal
Click for user's manual
Overview:
The
Leica SP2 is an advanced laser scanning confocal system with an
acousto-optical beam splitter (AOBS) to select/introduce most
excitation laser lines. There are nine excitation lines
available, spanning the
spectrum from blue to far red. The acousto-optical tunable
filters
(AOTF) make it possible to detect a wide range of emission wavelengths
with unlimited range on each of three detector channels.
Features:
- Inverted platform for imaging on slides or live
cell dishes
- Optical
zoom for increased magnification and resolution
- Sequential
scanning of fluorophores to minimize bleed-through and
cross-talk
- DIC polarizer/analyzer plus prisms
appropriate for each objective.
- Transmitted light detector for recording bright
field images.
- Galvo stage for Z-sectioning
- 3D
rendering and wizards for
FRAP and FRET
- Spectral
scanning for separation of fluorophores with similar ranges
(e.g. GFP and FITC)
- LCS
Leica confocal software with customizable user interface on WindowsXP
- Off-line
version available for download
Objectives:
- 10x / NA 0.4 dry
- 20x / 0.7 dry
- 40x / 1.2-0.75 oil
- 63x / 1.4-0.6 UV oil
- 100x / 1.40-0.7 oil
- 63x / 1.3 glycerol
Lasers
for excitation:
- near
UV --
405nm (DAPI,
CFP)
- Argon
-- 458nm (CFP), 476, 488 (GFP, FITC), 496, 514 (YFP)
- orange
HeNe -- 543nm (Cy3) and 594 (TexRed)
- red
HeNe -- 633nm (Cy5)
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Olympus DSU
"fixed cell" Spinning
Disk Confocal
Click for user's
manual or scale bar calibrations
Overview:
The Olympus "fixed cell" DSU is one of two Olympus spinning disk
confocal systems available in the facility. These systems can be used
for either widefield or confocal fluorescence capture, and since the
entire field of view is illuminated at once, z-stack and time lapse
capture can be faster than on the laser scanning confocals. The
fixed cell DSU features filters for a broad range of fluorophores, from
blue to far red, plus DIC optics.
Features:
- Image
capture through SlideBook software
- Inverted platform for imaging on slides or live
cell dishes
- Heated
stage platform available for live cell preps
- z-galvo stage for Z-sectioning
- 3D and 4D (volume over time) rendering through
SlideBook
- Neutral density filters available for long-term
light time lapse imaging
- DIC polarizer/analyzer plus appropriate prisms
for most objectives
Objectives
on the microscope:
- 10x / NA 0.3 dry
- 20x / 0.5 dry
- 40x / 1.0 oil
- 60x / 1.4 oil
- 100x / 1.35 oil
- 60x / 1.2 water
available (please ask):
- 2x /
0.06 dry
- 4x /
0.16 dry
- 40x /
0.6 dry LWD
- 150x /
1.45 oil
Excitation
and Emission filters:
- blue
(DAPI) -- ex: 387/11nm and em: 440/40
- greens
(FITC, GFP) -- ex: 485/20 and em: 525/30
- reds
(Cy3, Texas Red) -- ex: 560/25 and em: 607/36
- far
reds (Cy5) -- ex: 650/13 and em: 684/24
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Zeiss Axioplan
(upright, high resolution deconvolution microscope)
Overview:
This microscope is used
for high resolution immunofluorescence on fixed
or live specimens on slides. The high quality images can be further
enhanced by digital deconvolution on the analysis stations using
Openlab.
Features:
 High resolution, cooled CCD
camera system
designed for low-light scientific applications. The Photometrics
CoolSnap FxHQ Monochrome camera provides 12-bit digitation at
20mHz. The imaging array is 1392 x 1040 (6.45x 6.45 um pixels) .
- Improvision's
user friendly OpenLab software captures images on Mac OS X
- Upright
platform
- Automated
z-drive for z-stacks
- Dual
filter wheels with individual excitation filters for fluorescence
- Shuttered
epi-fluorescence and transmitted light paths
- Nomarksi
(DIC) imaging
Objectives
on the microscope:
- 10x / NA 0.25 NA
- 20x / 0.5 NA
- 32x / 0.4 NA long distance working lens
- 63x / 1.4 oil
- 100x / 1.3 NA oil
- (Also available: 1.2x, 4x, 25x Korr
oil/glycerol, 40x dry or oil)
Excitation
and Emission filters:
- Triple
emission filter: Dapi, Fitc, Texas red
- Single
emission filters: Texas Red, GFP, Fitc Long
- Quad
emission filter: Dapi, Fitc, Texas red, Cy5
- Dual
emission filter: CFP, YFP, FRET
Images obtained on
the Axioplan:
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Zeiss Axiovert 200M
(inverted widefield fluorescence)
The Axiovert 200M is the state of the art in inverted microscopy from
Zeiss. It is fully motorized and an absolute pleasure to use.This
microscope is used for high resolution immunofluorescence or Nomarksi
(DIC) images on fixed
or live specimens on slides. The high quality images can be further
enhanced by digital deconvolution on the analysis stations. Users may
capture their images using the ORCA ER digital camera for fluorescent
samples or the Zeiss Axiocam color digital camera for samples which
require transmitted light color imaging.
Features:
- High resolution, cooled CCD camera system
designed for low-light
scientific applications. The Hammatsu Orca ER firewire digital
monochrome
camera provides high resolution at 12 bits. The imaging array is
1344 x 1024 (6.45x 6.45 um pixels).
- Ultra-high
Resolution with the Zeiss
Axiocam digital color camera
- CCD
basic
resolution is
1300x1035;
- maximum file size per image is 3900x3030@3x14 bit
(color);
- Digitisation is14
bit/10MHz
- Pixel
size is 6.7 x 6.7 microns
- Improvision's
user friendly OpenLab software for image capture on Mac OS X
- Inverted
platform and multiple stages to support slides and dishes for live and
fixed samples
- Automated
z-drive for z-stacks
- Automated
xy stage for fine movement
- Automated
objective turret
- Automated
reflector turret with 5 positions for a multitude of fluorescent filters
- Automated
condensor turret
- Dual
filter wheels with individual excitation filters for fluorescence
- Nomarksi
(DIC) imaging
Objectives on the microscope:
- 10x / 0.5 NA
- 20x / 0.5 NA
- 40x / 1.3 NA oil
- 40x Korr
- 63x / 1.4 NA oil
- Also available:
- 1.25x / 0.035 NA
- 4x/0.10 NA
- 25x Korr oil/ glycerol
- 32x/0.4NA
- 40x / 0.6 NA
- 100x/ 1.4 NA oil
Excitation
and emission filters:
- Single
emission filter cubes: Dapi, CFP, Fitc, GFP, eGFP, YFP,Texas red, Cy3,
Cy5
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Zeiss Axiovert 100TV
(inverted widefield fluorescence for timelapse)
Overview: The
Axiovert 100TV is a versatile microscope. The user can simultaneously
obtain digital low light level fluorescent and video-enhanced contrast
Nomarksi (DIC) images of living cells or fixed samples. LIve cell
observation can be performed using an enclosed chamber or a Bioptechs
chamber. The enclosed chamber maintains physiological conditions for
extended time periods. Collect single or multiple site time lapse
images in 2 or 3D (Z-series).
Features:
- High resolution, cooled CCD camera system
designed for low-light
scientific applications. The Retiga EXi firewire digital monochrome
camera provides 10fps resolution at 12 bits. The imaging array is 1392
x 1040 (6.45x 6.45 um pixels)
- 3I's
user friendly Slidebook software is used for image capture on a PC
- Inverted
platform for fixed or live samples on slides, dishes, or plates
- Automated
z-drive for z-stacks
- Dual
filter wheels with individual excitation filters for fluorescence
- Shuttered
epi-fluorescent and transmitted light paths
- Nomarksi
DIC) imaging
- The
Microscope is enclosed in a chamber for CO2 and heat for live cells
Objectives on the microscope:
- 10x / 0.3 NA
- 20x Korr
- 40x / 1.3 NA oil
- 40x Korr
- 63x / 1.4 NA oil
- Also available:
- 1.25x / 0.035 NA
- 4x /0.10 NA
- 20x/ 0.5 NA
- 25x Korr oil/ glycerol
- 40x / 0.6 NA
- 100x 1.4 NA oil
Excitation
and emission filters:
- Single
emission filter: GFP
- Dual
emission filter: CFP, YFP
- Quad
emission filter: Dapi, Fitc, Texas red, Cy5
Images obtained from the Zeiss Axiovert 100TV:
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Zeiss Axioskop
(upright histology)
 Overview: This microscope is optimized
to meet the needs of those who want high resolution histological work.
Data can be captured with the digital camera and analyzed on an
analysis station.
Features:
- Ultra-high
Resolution with the Zeiss
Axiocam digital color camera
- CCD
basic
resolution is
1300x1035;
- maximum file size per image is 3900x3030@3x14 bit
(color);
- Digitisation is14
bit/10MHz
- Pixel
size is 6.7 x 6.7 microns
- Capture images with Improvision's user friendly OpenLab
software on Mac OS X
- Brightfield,
phase contrast and Nomarski (DIC)
- Upright
platform
Objectives
on the microscope:
- 1.25x /0.035 NA
- 4x / 0.10 NA
- 10x / 0.3 NA
- 20x / 0.5 NA
- 40x / 1.3 NA oil
- Also available:
- 25x / NA oil/ glycerol
- 40x / 0.6 NA or 40x Korr
- 63x / 1.4 NA oil
- 100x 1.4 NA oil
Images obtained from the Zeiss Axioskop:
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Olympus SZX-Zb12
Research Stereomicroscope
 Overview: The stereomicroscope is
built for imaging large specimens. The scope has fluorescent,
transmitted, and oblique illumination, 2 objectives, zoom focusing, and
a high resolution color camera.
Features:
- Long
working distance and wide zoom range (12:86:1)
- 10x
occulars
- External
fiber optic illuminator for oblique
lighting
- Fluorescence light: Stereo fluorescence with
100W mercury lamp, GFP
Longpass and Cy3 fluorescence filter set.
- Ultra-high
Resolution with the Zeiss
Axiocam digital color camera
- CCD
basic
resolution is
1300x1035;
- maximum file size per image is 3900x3030@3x14 bit
(color);
- Digitisation is14
bit/10MHz
- Pixel
size is 6.7 x 6.7 microns
- Improvision's user friendly software,
OpenLab, on Mac OSX
Objective:
- 0.5x
Plan fluorite PF (NA 0.055,
working distance 70mm) objective plus zoom
- 1.2x
PF2
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Olympus IX70 TIRF
Microscope (inverted)
Click for user's
manual or scale bar calibrations
 Overview:
This is a manual fluorescence microscope that is well suited to fixed
cell fluorescence imaging, live cell imaging with real-time pixel
intensity readouts, FRET experiments, ratiometric imaging (such as
fura-2 340/380), and checking transfection efficiency.
What
isTIRF? This microscope is currently our only TIRF
capable setup. Total Internal Reflection Fluorescence is used
to observe the events near the plasma membrane of cells, or the
fluorescence of a single molecule,  without
interference from possible background fluorescence. An evanescent
wave selectively excites fluorophores in a small region (less than
200 nm)
above the coverglass. This microscope features a single TIRF excitation
laser at 488nm. This laser can excite both green and some red
fluorophores, and combined with our emission beam splitter, the system
allows for one and two color TIRF experiments.
Movie of bovine
chromaffin cell exocytosis events
courtesy of Dr. Li Ma of the Philipson Lab and Dr. Vytas
Bindokas.
Features:
- Inverted platform for imaging on slides or live
cell dishes
- Retiga
camera with high sensitivity and resolution (6um pixels)
- Manual
filter changer with quad filter for multi-color fluorescence imaging
- DIC polarizer/analyzer plus appropriate prisms
for most objectives
- Heated stage platform available
- MetaMorph
for image capture in 2D, 3D or time lapse
- MetaFluor
software for image capture plus real-time fluorescence intensity or
ratiometric data readout/graphing
- TIRF excitation (488nm only, manual setting of
critical angle required)
- Beam splitter for simultaneous capture of green
and red fluorophores in both widefield and TIRF modes
- Fura-2
340/380 ratiometric imaging available
- CFP/YFP
and Chameleon FRET filter set available
Objectives
on the microscope:
- 10x / NA 0.3 dry
- 20x / 0.4 dry
- 40x / 0.6 dry LWD
- 60x / 1.45 oil (TIRFM)
- 100x / 1.3 oil
available (please ask):
- 2x /
0.06 dry
- 4x /
0.16 dry
- 40x /
1.2 oil UV (best for FURA)
- 100x / 1.4 oil (TIRFM)
- 150x / 1.45 oil (TIRFM)
Excitation and Emission filters:
- blue
(DAPI) single cube -- ex:
360-370 and em: 420LP
- blue
on quad -- ex:
380nm and em: 450/65
- CFP --
ex: 436/20 and em: 480/40
- greens
(FITC, GFP) single cube -- ex: 480/40 HQ and em: 510LP
- greens
on quad -- ex: 480/40
HQ and em: 535/30HQ
- YFP --
ex: 500/20 and em: 535/30HQ
- reds
(Cy3, Texas Red) single cube -- ex: 530-550 and em: 590LP
- reds
on quad -- ex: 620/60HQ and 682/22
- far
reds (Cy5) single cube -- ex: 620/60 HQ and em: 700/75 HQ
- Fura-2
ex: 340; 380 and em: 510/40
- Chameleons
2 ex: 440/20 and em: 535/35; 485/40
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Abbott Hall (AB-129) Microscopes
Leica SP5
Tandem Scanner
Spectral 2-Photon Confocal
 Overview:
This laser scanning confocal
system features software selectable
conventional or high-speed resonance scanner galvanometer system with
three (chilled, high sensitivity) internal PMT detectors (spectral),
two internal avalanche photodiodes, and two external (NDD) PMT
detectors. Six visible laser lines and a tunable NIR pulsed laser
(Spectra Physics Mai Tai broadband 710-990nm) provide
excitation. The system features turnkey operation and full software
control in addition to the features listed below:
Features:
- 3 chilled PMT fluorescence detectors
(digital spectral definition in 1 nm increments) plus one transmitted
light detector. 8-, 12-, or 16-bit output.
- 2 external non-descanned detectors; 2
internal APD detectors. (green/red and CFP/YFP filters
or CY7)
- Spectra Physics Mai Tai broadband
Ti:sapphire laser with 10W pump (tunable from 710 to 990 nm) (up to 2W
output at 810 nm)
- Dual scanning galvanometer mirrors (max
16,000 Hz scan rate) 25 images/sec at 512x512; strip scans to 333 fps
(4 channels). Standard scanner with beam park, bleaching,
imaging to 8k x 8k (64 megapixels) per channel.
- "SuperZ" galvo focusing stage,
±1.5 mm range, "instantaneous" XZY
scanning. Demonstrated
capability of three cell volumes per second (12 x 1-mm optical slices
each)
- AOTF control of visible laser lines, EOM
and ND filter attenuation of NIR
- Excitation and emission scanning
capability with linear unmixing
- Automatically optimized confocal pinhole
apertures
- Digital zoom to 40X
- Transmitted detector with LWD and oil
lenses and filters suited for SHG imaging
- FRET, deconvolution, and 3D software
options
- Custom NIR blocking microscope
jacket/incubator system with PID heater and CO2 capability
- Inverted platform for imaging of slides or
dishes
- LSM objective inverter device (allows
system to be used as if it was an upright microscope platform)
Excitation Laser Lines:
- Argon
-- 458nm (CFP), 476nm, 488nm (GFP, FITC), 514nm (YFP)
- orange
HeNe -- 561nm (Cy3, Texas Red)
- red
HeNe -- 633nm (Cy5)
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Olympus "live cell" DSU Spinning
Disk inverted confocal
Click for user's
manual or scale bar calibrations
Overview:
The Olympus "live cell" DSU is one of two Olympus spinning disk
confocal systems available in the facility. These systems can be used
for either widefield or confocal fluorescence capture, and since the
entire field of view is illuminated at once, z-stack and time lapse
capture can be faster than on the laser scanning confocals. The
fixed cell DSU features filters for a broad range of fluorophores, from
blue to far red, plus DIC optics.
Features:
- Back thinned EM-CCD Hamamatsu camera for high
speed/ high sensitivity imaging
- Image
capture through SlideBook software
- Inverted platform for imaging on slides or live
cell dishes
- Heated
stage platform and 5% CO2 chamber available for live cell preps
- Motorized xy-galvo stage for imaging multiple
areas per dish and automated tiling of large specimens
- z-galvo stage for Z-sectioning
- 3D and 4D (volume over time) rendering through
SlideBook
- 100% - 1.5% neutral density filters available
- Interchangable pinhole disks for many
magnifications (special request only)
- Dual cube viewing through the occulars
(blue/green, green/red and red/far red)
Objectives
on the microscope:
- 10x / NA 0.3 dry
- 10x / 0.4 water
- 20x / 0.7 water
- 40x / 1.15 water
- 60x / 1.2 water
- 100x / 1.45 oil
available (please ask):
- 2x /
0.06 dry
- 4x /
0.16 dry
- 40x /
0.6 dry LWD
- 150x /
1.45 oil
Excitation
and Emission filters:
- blue/violet
or blue (DAPI) ex: 350/50 or ex: 405/12 and em: 457/50
- CFP ex: 436/10 and em: 470/30
- greens
(FITC, Cy2) -- ex: 490/20 and em: 528/38
- greens
(GFP) -- ex: 480/25 and em: 525/40
- YFP
ex: 500/20 and em: 535/30
- reds
(TRITC, Cy3) -- ex: 555/28 and em: 617/73
- reds
(DsRed) ex: 565/25 and em: 620/60
- far
reds (Cy5) -- ex: 635/20 and em: 685/40
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Zeiss Axiovert135 (inverted widefield
fluorescence)
Click
for user's
manual
 Overview:
A manual microscope with simple controls for checking fluorescent
protein expression. Can do simple image capture (multi color
stills, single color time lapse).
Features:
- Manual
inverted microscope platform
- Image
capture with MicroMax camera and MicroManager software
Objectives:
- 20x / NA 0.5 dry
- 40x / 1.3 oil
- 63x / 1.4 oil
Fluorescence Excitation and Emission
Filters:
- blue (DAPI) --
ex: 365/50 em: 450/60
- green (GFP/FITC) --
ex: 450-490 em: 515-565
- red (rhodamine) --
ex: 546 em: 590LP
- far
red (Cy5) -- ex: 628/40 em: blank
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Leitz Diaplan (upright histology
and fluorescence microscope)
Overview: A small, upright
microscope with high-quality color camera for
histology.
Also features limited manual fluorescence, brightfield, DIC and
darkfield imaging.
Features:
- QImaging
MicroPublisher 3.3 RTV firewire camera
- Darkfield
and DIC filters (optimal
on 25x and 40x) plus
brightfield illumination
- 50/50
ocular and camera split view
- White
balance correction through the QImaging capture software suite
Objectives:
- 2.5x / NA 0.08
- 6.3 / 0.20
- 25 / 0.6
- 40 / 0.7
- 100 / 1.25 oil
Fluorescence Excitation / Emission Filters:
- blue
-- (DAPI)
- green
-- (GFP/FITC)
- red --
(rhodamine)
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Contact Information
Vytas
Bindokas, PhD
Facility Director
Technical Director, AB-129
773.702.4875
vbindoka@bsd.uchicago.edu
Shirley Bond, MA
Technical Director, KCBD 1250
773.834.2639
sbond@bsd.uchicago.edu
Christine Labno, PhD
Technical Director, KCBD 1250
773.834.9040
ccase@midway.uchicago.edu
Main Hospital Building
(Vytas)
Abbott Memorial Hall
Room AB-129
Knapp
Center
for Biomedical
Discovery
(Shirley & Christine)
First Floor, Room 1250
Core Support
Office of Shared
Research
Facilities (OSRF)
Cancer Research Center (UCCRC)
Intellectual and Developmental
Disabilities Research Center
(IDDRC)
Digestive Diseases Research
Core Center (DDRCC)
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