For SQUIRREL analysis of SIM images (Sup. Fig. 3 ), FluoCells prepared slide #2 (Invitrogen) with BPAE cells stained with Texas Red-X phalloidin and Alexa Fluor 488-tubulin was imaged on a Zeiss Elyra PS.1 system, using a 63x NA 1.4 objective with additional 1.6x magnification for SIM and widefield acquisition. For actin imaging, ‘Low SNR’ images were acquired with a 561 nm laser at 0.05 % laser power, using 100 ms exposure time, and 5 grid rotations. ‘High SNR’ images were acquired with a 561 nm laser at 5 % laser power, 100 ms exposure, 5 grid rotations. Widefield images were acquired with a 561 nm laser at 0.2 % laser power, 100 ms exposure time. SIM reconstructions were generated with the Zeiss Elyra Zen software using automatic settings. For microtubule imaging, raw SIM data was acquired with a 488 nm laser at 10 % laser power using 100 ms exposure time and 3 grid rotations. The SIM reconstruction was generated using FairSIM [19 (link)].
Fluocells prepared slide 2
Manufactured by Thermo Fisher Scientific
FluoCells prepared slide #2 is a laboratory equipment product designed to provide a standardized fluorescent cell sample for microscopy applications. The slide contains fixed and stained mammalian cells that exhibit specific fluorescent labeling patterns. This product is intended to serve as a reference sample for microscope setup, alignment, and performance evaluation.
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Lab products found in correlation
2 protocols using fluocells prepared slide 2
1
Super-Resolution Imaging of Cytoskeleton
2
Super-Resolution Imaging of Cytoskeleton
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For SQUIRREL analysis of SIM images (Sup. Fig. 3 ), FluoCells prepared slide #2 (Invitrogen) with BPAE cells stained with Texas Red-X phalloidin and Alexa Fluor 488-tubulin was imaged on a Zeiss Elyra PS.1 system, using a 63x NA 1.4 objective with additional 1.6x magnification for SIM and widefield acquisition. For actin imaging, ‘Low SNR’ images were acquired with a 561 nm laser at 0.05 % laser power, using 100 ms exposure time, and 5 grid rotations. ‘High SNR’ images were acquired with a 561 nm laser at 5 % laser power, 100 ms exposure, 5 grid rotations. Widefield images were acquired with a 561 nm laser at 0.2 % laser power, 100 ms exposure time. SIM reconstructions were generated with the Zeiss Elyra Zen software using automatic settings. For microtubule imaging, raw SIM data was acquired with a 488 nm laser at 10 % laser power using 100 ms exposure time and 3 grid rotations. The SIM reconstruction was generated using FairSIM [19 (link)].
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