Inverted Confocal Microscope

SLU is performing many live imaging experiments where there is a need to image deep into plants, microbes and insects, which is a big challenge. Multiphoton imaging is not a solution as pigments such as chlorophyll absorbs the light around 880-940nm needed for excitation. The main reason to buy Zeiss model LSM 880 with Airyscan is to avoid killing the microbes, or bursting plant cells. Furthermore to avoid photobleaching these live cell imaging experiments must be performed rapidly with high sensitivity. The inverted Zeiss LSM 880 Confocal microscope with Airyscan capitalizes on the scanning and optical sectioning capabilities of a confocal, it works on thicker samples such as tissue sections and whole organisms that require increased penetration depth, such as insect tissues, and infected plant materials. Therefore, SLU have the intention to buy 1 confocal microscopes from Zeiss model LSM 880 with Airyscan mounted on an inverted microscope. Read more under point IV.1 and VI.3.

SLU intends to buy one (1) confocal microscope from Zeiss model LSM 880 with Airyscan (incl. fast module) and PicoQuant FLIM/FCS module mounted on an inverted microscope.

SLU announces its intention and takes into account a standstill period of 10 days from the date of publication.

Förklaring:

The Zeiss LSM 880 is the only confocal microscope that allows SLU to acquire images with lower laser light intensity and higher resolution. It is a point scanning confocal that can also be used for super resolution using the pixel reassignment technology (Airyscan). This will give an isotropic increase in resolution down to 120 nm at 488 nm.

The main reason to buy Zeiss model LSM 880 with Airyscan is for deep scanning, whilst avoiding killing microbes, or bursting plant cells. The Zeiss LSM 880 with Airyscan capitalizes on the scanning and optical sectioning capabilities of a confocal, it works on thicker samples such as tissue sections and whole organisms,including insects that require increased penetration depth. It is also essential for monitoring structure-function relationships in bio-based materials. The Airyscan detector is essential for infected plant tissues where SLU can acquire images with high resolution at a greater depth than with other techniques. The 880 LSM Airyscan FAST is the only confocal microscope on the market that currently allows SLU to do this in a precise, fast and time resolved manner.

SLU have discussed the feasibility with other suppliers and all of whom SLU has contacted has referred to Zeiss as the only confocal microscope that can ensure that SLU gets desired results. As Zeiss has a patent on Airyscan, no other company (known for SLU) can provide SLU with this so vital function for imaging at SLU.

SLU has a need for deep live cell imaging from the bottom with an inverted microscope, in this case SLU has problems with depth penetration and sensitivity with SLU's current microscope and microscopes from supplier other then Zeiss.

The Airyscan detector is a revolutionary detector for Laser scanning microscopes (LSM). Utilizing a pinhole plane imaging concept, Airyscan allows for a simultaneous improvement in resolution and signal to noise by capitalizing on an innovative 32 channel GaAsP PMT array detector. Each detection channel functions as single very small pinhole of 0.2 AU to increase resolution (120 nm lateral, 350 nm axial) while the overall detector captures 1.25 AU and increases signal to noise by 48 times over traditional GaAsP based confocal systems. Building upon the Airyscan pinhole plane imaging concept, the optional Fast module for Airyscan allows a 4 fold increase in acquisition speed whilst retaining the resolution and signal to noise benefits. Unlike other super resolution techniques that require complicated biochemical labeling procedures or dye selections, the Airyscan is not more complicated than regular confocal microscopy. As universal as the LSM itself, it performs with any sample or fluorescent label that is excited by the available range of visible laser lines or multi-photon excitation. The improvements in sensitivity and resolution can even be seen with objectives of low magnification or numerical aperture (NA). Whilst some super resolution techniques require acquisition and processing times of minutes to hours, the Airyscan offers super resolution on the fly for fast sample screening.

The combination of live cell imaging with a 34 array detector and Airyscan FAST mean that SLU can perform new cutting edge research in insects, plant-microbe interactions and biobased materials. Frontline research on insect chemical ecology and neurophysiology requires imaging of live neural tissue, often of the central nervous system. Traditionally these experiments were performed with calcium ­sensitive dyes and ordinary fluorescent microscopes. However, modern techniques utilize transformed cells and animals that express, sometimes transiently, calcium flux markers, that permit penetration to greater depth and with enhanced resolution. To properly benefit from this, an LSM with sensitive and fast live­ cell imaging is critical. The Zeiss 880 LSM Airyscan offers the possibilities to use enhanced sensitive, rapid live cell imaging. In addition, the Airyscan FAST allow us to penetrate much greater depth in our preparations without compromising resolution. Scanning at depths greater than 100um are required for insect neural studies as well as plant-microbe interactions and biobased materials.

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