The Microscopy Imaging Core is located in the Research Core Facility on the sixth floor of the Biomedical Research Institute. This is a shared user facility that provides access to advanced microscopes as well as the expertise of dedicated imaging specialists. The microscopes are open to all members at LSUHSC-Shreveport and outside users. We are happy to assist you with instrument training, experimental design, image acquisition, and image analysis. We encourage you to contact us prior to beginning a new project.
All users must be trained by the RCF staff before using the microscopes. For training request, please contact Dr. Chaowei Shang.
- Are you new to fluorescent microscopy? Check out this paper to avoid errors and bias! http://jcb.rupress.org/content/218/5/1452
- Check out this new course on Bioimage analysis from iBiology! https://www.ibiology.org/online-biology-courses/bioimage-analysis-course/
|AxioObserver + Apotome||$15||$300||$525||$750|
|Leica TCS SP5 Confocal||$40||$800||$1500||$2000|
|Nikon A1R Confocal & Super-Resolution||$50||$1000||$1750||$2500|
|Zeiss LSM 510 Coherent Multiphoton||$50||$1000||$1750||$2500|
|Staff acquire and process image||$65/h for Nikon widefield and AxioObserver, $75/h for other scopes|
|AxioObserver + Apotome||$25||$450||$788||$1125|
|Leica TCS SP5 Confocal||$65||$1200||$2250||$3000|
|Nikon A1R Confocal & Super-Resolution||$75||$1500||$2625||$3750|
|Zeiss LSM 510 Coherent Multiphoton||$75||$1500||$2625||$3750|
|Staff acquire and process image||$75/h|
|AxioObserver + Apotome||$100|
|Leica TCS SP5 Confocal||$150|
|Nikon A1R Confocal & Super-Resolution||$200|
|Zeiss LSM 510 Coherent Multiphoton||$150|
- Quantitative colocalization
- Analyze Immunohistochemistry images in ImageJ
- Make Binary & intensity measurements on colocalized channels (Nikon-Nis-element)
- How to open a multichannel Z-stack image in Image J/FIJI
- How to convert a image to TIFF format in Image J/FIJI
- Create inset in an image for publication
|RCF offline 1||RCF offline 2||RCF offline 3||RCF offline 4|
|Computer||Dell OptiPlex 3010||HP Z440||Dell Precision 5820||Dell OptiPlex 5040|
LAS AF 2.7.3
|System||Windows 7, 64-bit||Windows 7, 64 bit||Windows 10, 64 bit||Windows 10, 64-bit|
|RAM||4 GB||64 GB||128GB||64 GB|
|Location||F 6-27||F 6-27||F6-13||F 6-13|
RCF Offline 1 and 2 are free to all LSU Health employees and outside users for image analysis purpose, and no reservation is needed. License dongles are required for full access to some of the software. Please contact Dr. Chaowei Shang for the dongles.
For most users, the use of RCF Offline 3 and 4 cost a minimal fee to maintain the yearly service contract with Bitplane Imaris. You get free remote support from the Imaris (Bitplane) team.
Biosafety for Live Sample Imaging
1. RCF Basic operating procedure for live sample imaging
All samples must be prepared in your lab following biosafety rules.
All live samples must be carried to the microscope room in a sealed secondary container with an absorbent material at the bottom of the container. Parafilm must be used to wrap the cell culture dish during sample transfer. Remember to take off the parafilm before you start the experiment for CO2 perfusion.
During sample transfer, you should keep one hand clean without gloves, and another hand with a glove on to hold samples.
Wipe the dish or chamber insert, the dish holder, and the microscope stage surface with 70% ethanol after each experiment.
Clean hands, no gloves. Area: Room doorknobs, light switches, flashlight, chairs, CO2 tanks, sliding doors of microscope box, computer, keyboard, mouse, temperature wrapping band, microscope system parts that are outside the microscope box and below the microscope stage.
With gloves. Area: Your samples, dish/chamber inserts, live cell dish holder, microscope stage.
There is a small plastic tray inside the microscope box to hold live cell dish holder. Place your gloves in the secondary container you brought with you. Never put gloves directly on the computer table.
If any medium spills out, wipe down the surface immediately with dry paper towel, then wipe down the dry surface with 70% ethanol 3 times. If medium spills out onto the microscope objectives, wipe the objectives with ethanol pads 3 times. Dispose of contaminated gloves immediately into the biohazard box in room F6-51. Inform RCF staff as soon as possible that a spill has occurred.
2. RCF Advanced operating procedure for live sample imaging
BSL (Biosafety level) Overview
BSL-1: Agents that do not cause disease in healthy human.
BSL-2: Agents of potential moderate biohazard to personnel and environment. (Lentivirus, HIV, HBV, all human cells which may contain pathogens, e.g. Hela cells).
BSL-3&4: Agents and microbes that can cause serious and lethal disease via the inhalation route (airborne pathogens).
BSL-3&4 live sample imaging is NOT ALLOWED in the microscopy core.
General Biosafety Guidelines (resource: Addgene)
Adenovirus: This virus can cause mild to severe respiratory disease in humans. Ethanol cleaning does not inactivate adenovirus; 10% bleach (0.5% sodium hypochlorite) should be used instead. Use of a replication incompetent adenoviral system (BSL-2) reduces risk but requires multiple rounds of plaque purification.
Adeno-associated virus (AAV): This virus is replication-incompetent and not known to cause disease in humans. If it is prepared using a helper plasmid rather than helper virus, it can often be handled at BSL-1. If prepared using a helper virus, it should be handled at BSL-2. Ethanol cleaning does not inactivate AAV; 10% bleach (0.5% sodium hypochlorite) should be used instead.
Lentivirus: Lentiviral systems are derived from HIV, but their organization across multiple plasmids and the deletion of many HIV proteins lowers the probability of generating replication-capable virus. These systems are handled at BSL-2/2+.
Retrovirus: Retroviruses are classified based on the cell types they infect. For retroviruses that do not infect human cells, BSL-1 may be appropriate; if human cells can be infected, BSL-2/2+ is appropriate.
Detailed resource: https://www.addgene.org/biosafety/
Basic Microscopy Information
Confocal microscopy - 39 things that can go wrong
Image processing: what's allowed and what's not
Leica Science Lab Microscopy Information
Extra Microscopy Information
Fluorescent dye exication and emission spectra look up
Scanning techniques and photobleaching
Tag and Turbo fluorescent proteins (Evrogen)
Fluorescent probes for super-resolution imaging: STED, FPALM, STORM