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Research Home
 User policy
Quick
reference guide
LSM
510 META
LSM 5 Live
Operation
manual
LSM
510 META
LSM-5-Live
Training materials
Internet resources
Online booking
Registration
form
System Administration
Contact us
Academic
Staff in-charge
Dr Liou Yih Cherng (ext 7711)
Confocal Microscopy Laboratory Staff
Tong Yan (ext 4757 and 7202)
Heng Mok Wei (Wang Muwei) (ext 2854 and 7202)
Location
Blk S1A, #03-03, |
The
Confocal Microscopy Laboratory,
located
at #03-03, Blk S1A, have two Carl Zeiss Laser
Scanning Microscopes (LSM) - LSM
510 Meta and LSM 5 Live
LSM
510 Meta
System
components:
- Laser
module (class 3b)
- Argon
multi-line gas laser (458, 477, 488, 514nm, 30.0mW)
- Helium
Neon (HeNe) gas laser (543nm, 1.2mW)
- HeNe
gas laser (633nm, 5.0mW)
- Diode
laser (405nm, 30.0mW)
- Microscope
- Axiovert
200M
- Objective:
- Filter
sets: 01,09,20
- LSM510
META Scan head
- Photomultiplier
tubes (PMT), META detector and PMT for DIC
- Beam
splitter cascade, emission filter
- Emission
filter
- PC
with LSM software (ver. 4.0 sp2 with Physiology)
Important
features/applications including:
- Z-stack:
generation of a Z-series that dissect through the specimen
to get 3D imaging.
- Co-localization
of proteins: multi-color imaging to detect the distribution
of various fluorescently-tagged proteins.
- Time
lapse: observation and record sample response under certain
treatment.
- Dynamic
study in live cells, i.e. Fluorescent Resonant Energy Transfer
(FRET), Fluorescent Recovery after Photo-bleaching (FRAP) and
Fluorescent Loss in Photo-bleaching (FLIP), etc.
- Meta
function: an advanced function to detect the full spectra of
multiple fluorophores and generate references of distinct spectral
signature to solve problem of emission cross-talk and tissue
background auto fluorescence.
LSM
5 Live
As
a ultra-fast scan confocal microscope, the LSM 5 live has accomplished
paradigm shifts from conventional LSM point scan to line scan
by 1) the complex beam shaper which converts the collimated
circular Gaussian laser beam into light with a rectangular cross-section
(line) and then focuses it on the specimen, 2) the color-independent AchroGate
beam splitter, which has virtually 100 % excitation efficiency
and a highly effective emission yield, and 3) the ultra-fast line
CCD detector allows parallel imaging of 512 pixels with quantum
efficiency of more than 75 %. It has been particularly designed
for studies on living specimens, e.g. fast Ca2+ release, tracking
mitochondrial movement, fast cell movement, etc.
LSM
5 Live can scan up to 120 confocal frames per sec with image
quality 512 x 512 pixels. Line scanning speed is up to 60,000
lines pr. sec. All scan combinations of line, X, Y, Z
and time are possible. Available laser lines are 405 nm, 488
nm and 561 nm. A CO2 incubator with temperature and humidity
control is available for live cell imaging. Objectives filter
sets and incubation control processed by a docking TFT touch
screen has been stationed.
System
components:
- Laser
module LIVE 230V (class 3B):
- Violet
- Laser diode, 405nm (50mW)
- Blue
- Laser diode, 488 nm (100mW)
- Green
- Laser diode, 561 nm (40mW)
- Microscope
- Inverted, Axio Observer.Z1
- Filter
sets: 49 DAPI, 38 Endow GFP and 43 CY3 shift free
- Objective:
- EC
Plan-Neofluar 10x/0.3
- EC
Plan-Neofluar 40x/0.75
- plan-Apochromat 63x/1.40 Oil DIC (with DIC sliders)
- LSM5
Live Scan head (Max. 120 fps at 512 x 512 pixels)
- One
channel
- line
CCD (1k x 1k pixels)
- Emission
filter
- CO2
incubator
- LCD
docking TFT touch screen, PC and monitor
- Software
- Zen 2007
Important
application including:
- Tracking
of morphological change, organelles, cell movement, microorganism
and mediators (e.g. Ca2+)
- Photo
manipulation: Photo-bleaching (e.g. FRAP, FLIP), Photo-activation & Photo-conversion
(e.g. PA-GFP, Keade, Dronpa)
Please
be responsible and follow usage policy closely to ensure effective
operation of the facility. |
