MicroBites
Issue
March 2010
The latest news from the Biomedical Imaging Research Unit
4
New Olympus FV1000
Confocal Microscope
In this issue
New appointment
New addition to the family
- Olympus Confocal
Microscope
New capabilities to the
existing microscopes
- TEM
- Nikon TE2000E
- Zeiss Axioplan2
- Zeiss LSM 710
- Leica SP2
- Olympus Live Cell
CLSM
- Heating and
Chamlide chamber
Upcoming seminars
New appointment
The new upright Olympus FluoViewTM FV1000 Confocal Microscope was
purchased to provide an additional system for imaging slides.
The FV1000 is a high resolution, high speed, highly sensitive confocal
system which has:
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Lin-Chien has recently
joined BIRU as a new
staff member on a parttime basis. She will train
users on microscopes,
including the Zeiss LSM
710 confocal microscope,
the
Nikon
TE2000E
inverted
fluorescence
microscope and the Zeiss
Axioplan2 fluorescence
microscope. Lin-Chien will
also provide assistance
with image processing
software such as Amira
and Image Pro Plus
3D. Her email address
is
lc.huang@auckland.
ac.nz and her working
hours are from Monday to
Wednesday lunchtime.
two detectors with tunable bandwidth
one standard filter based detector
transmitted light detector for brightfield or DIC imaging
laser wavelengths: 405nm, 440nm, 473nm, 559 and, 635nm
objectives: 10x/0.4, 20x/0.8oil, 40x/1.25oil, 60x/1.35oil,100x/1.4oil.
The operating software is an easy-to-use menu driven interface which makes
both the imaging of multi-labelled specimens and post capture processing
quick and easy.
For more information or training, please contact Hilary at [email protected].
Click here to visit our BIRU website for more details on the specifications of this system.
Upcoming seminars: FMHS 501-505
Image Pro Plus 3D Suite
Lin-Chien Huang, BIRU, FMHS.
1 - 2pm, Tuesday 16th March
High-Resolution Immunofluorescence and Correlative
Microscopy: Applications in Cell and Tissue Biology
Electron
Professor John Robinson, Professor of Physiology and Cell Biology, The Ohio State
University
3 - 4pm, Friday 26th March
http://www.fmhs.auckland.ac.nz/sms/biru
Editor: Lin-Chien Huang
[email protected]
MicroBites Issues 4: March 2010
For Nikon TE2000E Inverted Microscope
For Zeiss LSM 710
The addition of a differential interference
contrast (DIC) slider enables this microscope to perform polarised light microscopy.
This is a contrast-enhancing technique for
birefringent structure that is often used in
combination with special stains such as
Picro-sirius red (for collagen) or Congo Red (for
amyloid).
The installation of Visual Macro Editor on the
Zeiss LSM 710 provides our researchers with
a graphical interface for creating and editing
macros. This allows the user to programme
routine image acquisitions, and package
multiple time series without needing advanced
programming experience.
Polarised light image of Picro-sirius red showing collagen
fibres in kidney tissue. Note that some fibres appear green and
others orange/red.
A new Plan Fluor 40x oil immersion objective
lens with NA 1.3 and working distance 0.2mm is
now available on this microscope. This objective
lens allows imaging of fluorescently labelled
cells in glass bottom dishes, or plates, at higher
resolution and with better performance than the
dry 40x objective lens.
Useful functions include:
Scanning automation, multi-point imaging
over time.
For Zeiss Axioplan2 Upright Microscope
The addition of an Olympus DP72 12.8 megapixel cooled
colour camera to this fluorescence microscope provides the
ability to take colour images.
The camera is suitable for those who prefer to image in “true”
colour and do not require the higher sensitivity of the cooled
monochrome camera. Polarised light microscopy, which requires
a colour camera, is now also possible on this microscope.
DP72
Camera
The MetaMorph software, which controls this system, has
been upgraded to version 7.6.1. New functions, such as 3D/4D
rendering and measurement, and a Micronuclei Application
Module, to detect micronuclei in mononucleate and binucleate
cells, are now available.
http://www.fmhs.auckland.ac.nz/sms/biru
Editor: Lin-Chien Huang
[email protected]
MicroBites Issues 4: March 2010
For Leica SP2 CLSM
The microscope is now mounted on an antivibration table. The microscope is now isolated
from vibration that may interfere with acquiring
perfect images.
Warner Instruments Heating Chamber
This heating chamber is suitable for 35mm
Petri dishes and can also be used with the new
Chamlide chamber. There are microscope
stage adapters for the following microscopes:
Zeiss Axioplan2 fluorescence microscope, Leica
TCS SP2 CLSM and the Zeiss LSM 710. This is
an open-dish chamber and is suitable for shortterm live cell imaging only since carbon dioxide
supply is not possible. The chamber can be used
with dipping objective lenses on either the Zeiss
Axioplan2 or Leica TCS SP2. It can also be used
on the Zeiss LSM 710 but a glass bottom dish is
required for optimum imaging.
For Olympus Live-Cell Imaging CLSM
A new filter cube, labelled as WIB, has been now
added to the filter wheel of the Olympus Live-cell
Imaging CLSM. Specifications are: Excitation
filter BP460-495, Emission filter BA510-IF,
Dichromatic mirror DM505. The advantage
of this long pass barrier filter includes: (1)
improved visualisation of green fluorophores
such as FITC and Alexa 488 through collecting
more signal; (2) ability to check the occurence
of bleedthrough for red-green double labelling;
(3) ability to identify autofluorescence due to
lipofuscin.
Chamlide Chamber for Live Cell Imaging: An
Alternative to the Glass-bottom Petridish
This chamber has a magnetic body to
provide a leak-free space for a 25mm circular
coverslip and can fit in any 35mm Petri dish
holder. There are chambers for both inverted
and upright microscopes. These chambers
can also be used, if necessary, to confine
specimens that are not easy to mount on
slides. Circular coverslips
which can be
sterilised and used to grow cells in multiwell
plates available for trial.
(This image is from Kyodo International Inc website.)
http://www.fmhs.auckland.ac.nz/sms/biru
Editor: Lin-Chien Huang
[email protected]
MicroBites Issues 4: March 2010
For Transmission Electron Microscopy
Glow Discharge Unit
Vacuum Embedding Oven
Grids coated with a carbon support film are
routinely used in TEM for negative staining
preparations, a common technique used for
looking at nanoparticles, nanocapsules and
viruses. However these support films tend to
be hydrophobic which inhibits the spreading of
particles onto the coated grid.
When processing dense tissue for TEM,
adequate infiltration of the embedding resin into
the sample can be a problem. Poorly infiltrated
tissue is difficult to cut and liable to fall apart in
the electron beam.
Glow discharge treatment with air makes the
filmed grids hydrophilic and negatively charged.
This treatment facilities optimal absorption of
samples onto the grid.
Embedding in an oven under vacuum improves
the infiltration of the resin into the sample and
facilitates the removal of air bubbles.
Minifridge
The Boekel MiniFridge II cold block incubator
is a convenient and reliable way to process EM
samples at temperatures from ambient down to
4OC.
The “dry” cold of the MiniFridge II makes it an
ideal substitute for messy ice buckets.
If you have any feedback or suggestions, please email Lin-Chien at [email protected]
http://www.fmhs.auckland.ac.nz/sms/biru
Editor: Lin-Chien Huang
[email protected]