Zeiss 880 Laser Scanning Confocal microscope with FCS, FLIM, and AiryScan
Location: SR C 17

Signup to use microscope. 

Instructions startup/shutdown/general imaging:  [Word doc]   [PDF]      How to do tiling.  

Instructions AiryScan:  [Webpage]  

This microscope is most commonly used for standard fluorescence confocal. It also has other imaging modes, a few listed here:

• reflection, IRM
• transmitted light
• automated stage for tiling large areas or repeatably imaging multiple positions
• live and fixed samples
• FLIM (time domain, excitation with pulsed 440 nm laser)
• FCS and FCCS (continuous wavelength excitation)

A few of the specifications:

• Zeiss AxioObserver inverted stand
• environmental chamber for temperature and stage insert for CO₂ control
• 10X air N.A. 0.3 -
  For locating samples by eye, not recommended for confocal (we know you will use it anyhow...)
• 20X air Plan-Apochromat N.A. 0.8 M27
  For thinner samples, cell culture or thin cut tissue.
• 25x/0.8 LD LCI Plan-Apochromat Imm Corr DIC M27 for oil, water, silicone oil or glycerine immersion (D=0-0.17mm) (WD=0.57mm at D=0.17mm)
  Long working distance immersion lens great for cleared tissues.
• 40X water N.A. 1.1 for standard imaging
• 40X water N.A. 1.2 for FCS
• 63X oil 1.40
• 100X oil N.A. 1.46 Use only for Airyscan and other specialty work.
  NOT FOR ROUTINE CONFOCAL Instead, use the 63X with a zoom set or large pixel size image.
• Excitation lasers: 405, 440 (pulsed for time domain FLIM), 458, 488, 514, 561, 594, 633 nm
• Detectors, path 1:
  * PMT, 410 to 750 nm
  * 32 channel spectral, 409 to 695 nm (example of true spectral detection here)
  * redshifted GaAsP, up to 750 nm
  * transmitted detector; use with Omega Optical 750SP filter in the condenser to block IR light from Definite Focus (see similar configuration on Zeiss 800 here).
• Detectors, path 2:
  * 2 GaAsP detectors ("BIG" detectors) with standard filter blocks for green/red and CFP/YFP
• Detector, path 3:
  * AiryScan (example with gamma tubulin)
• Automated Stage (tiling, repeated multiple positions)
• Definite Focus to lock focus for time lapse -- 750SP Omega Optical filter in condenser to prevent interference with transmitted light detector
• Windows 7 Zen Black

To adjust parfocaility using the touchpad:
Home > settings > focus > parfocality > adjustment

Other links

• How to open CZI files on your own computer:
  1. https://www.zeiss.com/microscopy/us/products/microscope-software/zen-lite.html
  2. ImageJ with BioFormats
• Multifield timelapse setup instructions
• Explanation of noise, scan speed, and filtering for practical image quality.
• AiryScan example with gamma tubulin
• A method for labeling structure overlaps, here at high resolution using AiryScan data
• Example of FLIM
• Live cell mitochondria imaging with this microscope

Examples of true simultaneous transmitted light imaging with fluorescence:



Click here for original size image.

Old website here for reference: indexpre20180701.html