Zeiss LSM 880 with Airyscan

Principle of Airyscanning


The Airyscan detector consists of 32 elements based on GaAsP PMT technology with each element detecting the emission signal of a focal spot much smaller than the typical 1 Airy volume. This detection scheme together with adapted deconvolution algorithms allows resolving fluorescently labelled structures to a much higher degree. In addition the detector can be used as an additional confocal detector and in combination with other detectors of the system. The achievable scan speed is from a range of 13 fps to 430 fps for 512 x 512 pixel resolution.

 

Illumination Source

 

A. Halogen illuminator for transmitted light- HAL 100 illuminator with quartz collector with 12V 110W Halogen rectangular filament
B. Metal Halide illumination- Illuminator HXP 120V inclusive built in power supply, lamp module and infrared filter
C. Fluorescence light source with motorized brightness switch and integrated motorized shutter, that works stepper motor controlled, fast and vibration free.
D. Epi-fluorescence equipment- Filter set 49 DAPI shift free, Filter set 43 Cy 3 d=25 shift free, Filter set 38 Endow GFP shift free, Filter set 46 YFP shift free- Ex BP 500/20; Em BP 535/30, Filter set 47 CFP shift free Ex BP 436/20; Em BP 480/40, Filter set 50 Cy5 shift free- Ex BP 640/30; Em BP 690/50

 

Objectives

 

• 10X/0.45 M27 Plan Apochromat (WD= 2.1 mm)
• 20X/0.4 Corr M27 LD Plan Neofluar (D= 0- 1.5 mm)
• 20X/0.8 M27 Plan Apochromat (WD= 0.55 mm)
• 40X/1.3 oil DIC M27 Plan Apochromat (WD= 0.2 mm) (UV)VIS-IR
• 40X/1.2 W Corr M27 C- Apochromat (D= 0.14- 0.19 mm) (UV)VIS-IR Selected for FCS
• 63X/1.4 oil DIC M27 Plan Apochromat (WD= 0.19 mm)
• 100X/1.4 oil DIC M27 Plan Apochromat (WD= 0.17 mm)

Color Cooled CCD Camera: Axiocam MRC: High resolution microscopy camera, basic resolution 1.4 Mega pixels, pixel size 6.45 x 6.45 µm
LSM 880 Spectral confocal scan head with 2 MA-PMT + 32 GaAsP including lasers BLD 405 nm, Ar 458/488/514 nm, HeNe 543 nm, HeNe 594 nm & HeNe 633 nm with ZEN software

 

Laser modules:

 

Diode 405 nm
Argon Multiline 458/488/514                                                                                                                                                                    HeNe 543 nm                                                                                                                                                                                                  HeNe 594 nm                                                                                                                                                                                                    HeNe 633 nm

 

Control Computer and Monitor

 

User PC advanced for LSM 7 systems, HP Z820 Workstation, CPU 1X Intel Xeon E52643, memory 16GB included, max 128 GB RAM. Softwares: The latest ZEN 2 with enhanced functionalities has been provided (e.g. Advanced 2D/3D/4D/5D/6D imaging including deconvolution, Spectral unmixing with emission finger printing to eliminate bleed through, photoactivation conversion with real ROI in a single field and many more. ZEN module FRAP, ZEN module FRET and ZEN module Experiment Designer hardware. All accessories for Fluorescence Correlation Spectroscopy (FCS)/FCCS module with internal 32-Array GaAsP is provided.

 

 

Zeiss LSM 510 Meta

Zeiss LSM 880

Zeiss LSM 880 in Airyscan mode

Cos7 cells treated with anti-Tom20 tagged with AF532 and anti-tubulin tagged with OG488, images taken in Zeiss LSM 510 Meta, LSM 880 and LSM 880 in Airyscan mode for comparing the lateral resolution capacities of the microscopes; Courtesy, Bioimaging facility, IISc.

Image credits: Numbering from left to right

Image 1: Section of thorax muscles of Drosophila stained with phalloidin TRITC and MitoGFP, Courtesy to Prasanna Katti, Upendra Nongthomba Lab

Image 2: Drosophila embryo with PI and Twist-tagged to FITC, Courtesy to Saroj Jawkar, Upendra Nongthomba Lab

Image 3: HeLa Kyoto mitotic Cells (dividing) with DAPI (Blue) excited by 405nm , fragmented Tubulin ( green) excited with 488nm and ACA Centromeres ( red) excited by 543nm, Courtesy to Carl Zeiss, India

Image 4: Cardiomyocytes stained with eGFP, cTnI (red) and DNA stained with DAPI, Courtesy to Divya S., Seshagiri PB Lab

Image 5: Arabidopsis root tip stained with PI, Courtesy to Shyam., Utpal Nath Lab

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