simpleISM—A straight forward guide to upgrade from confocal to ISM

Resolution in a confocal laser scanning microscopes (CLSM) can be improved if the pinhole is closed. But closing the pinhole will deteriorate the signal to noise ratio (SNR). A simple technique to improve the SNR while keeping the resolution same by upgrading the system to an image scanning microsco...

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Detalles Bibliográficos
Autores principales: Goswami, Monalisa, Lachmann, René, Kretschmer, Robert, Heintzmann, Rainer
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2022
Materias:
Lab Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9794089/
https://www.ncbi.nlm.nih.gov/pubmed/36574395
http://dx.doi.org/10.1371/journal.pone.0279378
Descripción
Sumario:Resolution in a confocal laser scanning microscopes (CLSM) can be improved if the pinhole is closed. But closing the pinhole will deteriorate the signal to noise ratio (SNR). A simple technique to improve the SNR while keeping the resolution same by upgrading the system to an image scanning microscope. In this paper, we explain in detail, based on an Olympus Fluoview 300 system, how a scanning microscope can be upgraded into an image scanning microscope (ISM) using a simple camera-based detector and an Arduino Due providing a galvo driving and camera synchronization signals. We could confirm a resolution improvement as well as superconcentration and made the interesting observation of a reduced influence of laser fluctuations.

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