Near infrared laser provoques focus variation in a confocal microscope

Remy Avila; Melissa Matrecitos-Avila; Reinher Pimentel-Domínguez; Elisa Tamariz; Pablo Loza-Alvarez; Edén J. Parra-Fuentes

doi:10.1117/12.3022743

20 June 2024 Near infrared laser provoques focus variation in a confocal microscope

Remy Avila, Melissa Matrecitos-Avila, Reinher Pimentel-Domínguez, Elisa Tamariz, Pablo Loza-Alvarez, Edén J. Parra-Fuentes

Author Affiliations +

Proceedings Volume PC13006, Biomedical Spectroscopy, Microscopy, and Imaging III; PC130060B (2024) https://doi.org/10.1117/12.3022743
Event: SPIE Photonics Europe, 2024, Strasbourg, France

Abstract

Precision and stability in confocal microscopy are vital for maintaining image clarity and preventing misinterpretation of radiometric fluctuations as changes in fluorescence intensity within the sample. This study discusses a focus variation induced by an 810-nm continuous wave laser integrated into the optical path of an inverted confocal microscope with a 60x oil immersion objective. Activation or deactivation of the laser leads to a drift in the focus position towards lower or higher values of the vertical coordinate z, respectively. The maximum observed drift is 2.25 μm, occurring with a 40 mW laser power at the sample over a 600-second exposure time. The temporal behavior of the focus position follows exponential curves resembling temperature fluctuations associated with a heat source. Our analysis strongly indicates that the focus drift is attributable to the heating of the immersion oil.

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Citation Download Citation

Remy Avila, Melissa Matrecitos-Avila, Reinher Pimentel-Domínguez, Elisa Tamariz, Pablo Loza-Alvarez, and Edén J. Parra-Fuentes "Near infrared laser provoques focus variation in a confocal microscope", Proc. SPIE PC13006, Biomedical Spectroscopy, Microscopy, and Imaging III, PC130060B (20 June 2024); https://doi.org/10.1117/12.3022743

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