Noise reduction technique for fluorescence imaging in multifocal multiphoton microscopy

Inkeon Ryu; Eunwoo Park; Eunbin Kim; Daekeun Kim

doi:10.1117/12.2624203

11 January 2022 Noise reduction technique for fluorescence imaging in multifocal multiphoton microscopy

Inkeon Ryu, Eunwoo Park, Eunbin Kim, Daekeun Kim

Author Affiliations +

Proceedings Volume 12159, SPIE Advanced Biophotonics Conference (SPIE ABC 2021); 1215902 (2022) https://doi.org/10.1117/12.2624203
Event: SPIE Advanced Biophotonics Conference (SPIE ABC 2021), 2021, Busan, Republic of Korea

Abstract

Multifocal multiphoton microscopy (MMM) by forming multifocal created with a lenslet array enables high-speed imaging than single-focal multiphoton microscopy. However, most multifocal multiphoton microscopy has a problem of degrading image quality compared to single-focal microscopes. In order to solve this problem, it is necessary to equalize the intensity of the multifocal incident light. The fluorescence signal emitted from the fluorescence material is determined by the intensity of the incident light, but compared to the single-focal multiphoton microscope, the incident light passing through the lenslet array not only causes problems in acquiring fluorescence images but also limits the field of view (FOV). This problem was overcome by using a specially designed beam shaper to equalize the multifocal incident light intensity. Another problem with MMM is that typical biopsy samples are opaque, so light scattering occurs when light is irradiated onto the sample. This is a cause of the noise that degrades image quality by generating crosstalk in which fluorescence signals emitted from each focus deviate from the focal plane and interfere with each other. For the minimization of crosstalk, we set up the algorithm technology to correct the sensitivity non-uniformity between channels of multi-channel photomultiplier tubes (PMT). And to minimize crosstalk between PMT circuits, an independent short-channel photon discriminator was designed to expand the number of focal points of the MMM system. Among the problems of MMM, two techniques were introduced, and it was confirmed that the noise of the fluorescence image was reduced. ¬

Citation Download Citation

Inkeon Ryu, Eunwoo Park, Eunbin Kim, and Daekeun Kim "Noise reduction technique for fluorescence imaging in multifocal multiphoton microscopy", Proc. SPIE 12159, SPIE Advanced Biophotonics Conference (SPIE ABC 2021), 1215902 (11 January 2022); https://doi.org/10.1117/12.2624203

ACCESS THE FULL ARTICLE

INSTITUTIONAL
Select your institution to access the SPIE Digital Library.

SELECT YOUR INSTITUTION

PERSONAL
Sign in with your SPIE account to access your personal subscriptions or to use specific features such as save to my library, sign up for alerts, save searches, etc.

PERSONAL SIGN IN

No SPIE Account? Create one

;

PURCHASE THIS CONTENT

SUBSCRIBE TO DIGITAL LIBRARY

50 downloads per 1-year subscription

Members: $195

Non-members: $335 ADD TO CART

25 downloads per 1 - year subscription

Members: $145

Non-members: $250 ADD TO CART

PURCHASE SINGLE ARTICLE

Includes PDF, HTML & Video, when available

Members: $17.00

Non-members: $21.00 ADD TO CART

PROCEEDINGS
4 PAGES

DOWNLOAD PAPER SAVE TO MY LIBRARY

GET CITATION

RIGHTS & PERMISSIONS

Get copyright permission Get copyright permission on Copyright Marketplace

KEYWORDS

Luminescence

Multiphoton microscopy

Image quality

Microscopes

Image acquisition

Beam shaping

Denoising

Show All Keywords

Keywords/Phrases

Search In:

Publication Years