SSBD:database

Project ID

530-Isobe-SpheroidCellPhotoconversion

Project SID

530

Title

x-y cross-sectional images of EpH4-spheroids expressing nucleus-localized PSmOrange2 before and after photostimulation of target cells

Description

Significance Three-dimensional (3D) two-photon patterned illumination using a combination of computer-generated holography (CGH) and wide-field temporal focusing (TF) has emerged as a highly effective approach for photostimulation. However, even though the axial full-width at half-maximum of a single-spot by TF is smaller than the single-cell size of 10 μm, the axial resolution of 3D multispot patterns produced by CGH with TF is lower than the single-cell resolution as a result of interference among multispots. Aim We aim to achieve 3D two-photon patterned illumination with single-cell resolution by combining CGH with time-multiplexed multiline temporal focusing (TM-ML-TF), which is implemented by adding an echelle grating at a position conjugate to the focal plane of the TF-CGH system. Approach We measure the 3D two-photon fluorescence distributions generated by the TF-CGH and TM-ML-TF-CGH systems. Results In TM-ML-TF, the crosstalk artifacts between the target spots in two proximal planes with an axial distance of 20 μm were suppressed from 81% to 15% compared with those in TF. We successfully achieved the photoconversion of 3D target cells in spheroids with single-cell resolution. Conclusions TM-ML-TF successfully suppresses the interference among multispots, enabling the TM-ML-TF-CGH system to provide precise 3D patterned illumination with single-cell resolution.

Submission Date

2026-08-03

Opened Date

-

Release Date

2026-10-01

Update Date

-

Kind

License

Project URL

-

Project DOI

-

Metadata Version

-

Template Version

3.0.3

Funding Information

This work was partially supported by the JST CREST (Grant Nos. JPMJCR1851, JPMJCR1921, JPMJCR24B2, and JPMJCR2433), JST PRESTO (Grant No. JPMJPR14F2), AMED (Grant Nos. JP19dm0207090 and JP21bm0704060), MEXT/JSPS KAKENHI (Grant No. JP18H04750 “Resonance Bio”), and the Nakatani Foundation for Advancement of Measuring Technologies in Biomedical Engineering grant program.

Total

4 files / 119.7 MB

Image datasets

zipped 2 files / 39.6 MB, raw image 2 files / 80.0 MB, total 4 files / 119.7 MB

Quantitative datasets

zipped 0 files / 0 bytes, raw bdml 0 files / 0 bytes, total 0 files / 0 bytes

Organism

Strain

-

Cell

Cell line

MeSH

Anatomical Entity

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UBERON

Biological Process

Cellular Component

Molecular Function

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Medical Subject Headings

Method involved in biological imaging