Fluorescence and Brightfield Microscopy

A range of modern microscopes is available for routine research use.

Each is fitted with a high quality digital camera for image acquisition.

Microscopes

 

Image Viewing, Processing and Analysis of Fluorescence Images

Images acquired using the Fluorescence and Brightfield microscopes within Flinders Microscopy will usually be in .tif fomat (TIFF files). This is the recommended format for microscopy images. TIFF files can be opened via picture viewing software on your own computer.
ImageJ (http://rsb.info.nih.gov/ij) and Cell Profiler (www.cellprofiler.org) are two freely downloadable software that can be used to carry out a wide range of image processing and analysis functions.
Adobe Photoshop (www.adobe.com/products/photoshop.html) can be used to ‘false colour’ images, merge images and adjust brightness and contrast with a high degree of precision.

Users

Upon successfully completed training, users are given access to the facility and the respective instruments for which they have been trained. All equipment use must be booked, and user fees apply.
User fees are kept to a minimum realistic rate. Funds from user fees go back into the Facility to cover repairs, parts replacement and equipment upgrades.
Brightfield and fluorescence microscope user fee charges are invoiced to laboratory supervisors bi-annually.
Please refer to the current table of User Fees (PDF 13KB) for more information.

Booking Equipment

All usage of microscopes and associated equipment must be booked by using the online Calcium Booking Calendar.

The calendars may be viewed by everyone. However, the ability to book specific equipment will only be enabled after successfully completed training with Flinders Microscopy Management Personnel.

Useful Resources and Link

Fluorescence imaging principles and virtual instrument / simulator tutorials:

Software to analyse microscopic images

Tutorials on the basics of fluorescence, excitation and emission – Life Technologies

 

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OLYMPUS BX50 FLUORESCENCE MICROSCOPE (upright microscope)

This microscope is suitable for imaging most common blue, green, red and far-red fluorophores - refer to -  and has 4x, 10x, 20x and 40x dry objective lenses (40x to 400x total magnification).  Illumination is by a LED light source. Regular transmitted light imaging is also possible. Most first-time users who require fluorescence microscopy are trained on this upright microscope unless there is a particular reason for using another microscope.
Equipped with a Photometrics CoolSNAP cooled CCD camera and MicroManager Imaging Software.

OLYMPUS AX70 FLUORESCENCE MICROSCOPE (upright microscope)

This microscope is suitable for imaging most common blue, green, red and far-red fluorophores (but has some minor differences in the specifications of the filter cubes compared to the Fluorescence BX50 microscope, refer to .
This microscope has 40x and 100x oil immersion objective lenses as well as 4x, 10x, 20x and 40x dry objective lenses (40x to 1000x total magnification). Illumination is by a LED light source.
Regular transmitted light imaging and DIC (differential interference contrast) imaging is available.
A live video camera is also available which may help in locating far-red fluorophore labelling, which is invisible to the human eye unless it is exceedingly bright.
Equipped with a Hamamatsu Orca cooled CCD camera and MicroManager Imaging Software.

OLYMPUS IX71 FLUORESCENCE & CONSTRAST MICROSCOPE (inverted microscope)

This microscope is designed for fluorescence or brightfield / contrast imaging of live or fixed cells in tissue culture flasks/plates.It is suitable for imaging common blue, green, red and far-red fluorophores (see ) as well as phase contrast and relief contrast imaging.  It is equipped with 10, 20x and 40x dry objective lenses (100x to 400x total magnification). Illumination is by a LED light source.
Equipped with a Soft Imaging Systems F-View CCD camera and AnalySIS Imaging Software.

  

 

 

 

 

 

 

 

 

 

BRIGHTFIELD OLYMPUS BX50 MICROSCOPE (upright microscope)

This microscope is suitable for mounted slides where fluorescence is not required.  It is equipped with a digital colour camera, and 2x, 4x, 10x, 20x and 40x dry objective lenses and a 100x oil immersion objective lens.  Multiple condensers are used on this microscope depending on the chosen lens; training therefore includes how to change condensers and set the correct condenser height and field diaphragm aperture to achieve optimal illumination (“Koehler Illumination”).

Equipped with a Q-Imaging colour CCD camera and Q-Capture software.

 

SZX10 STEREO MICROSCOPE/MACROSCOPE (dissecting scope with reflected light illumination)

This microscope is suitable for low magnification (6.3x to 63x total magnification) imaging with reflected light, and may be used for a range of specimens.Equipped with an Olympus DP20-5E digital imaging system.


 

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