Evidence map›Paper›PMID 42810836›Full record

ReviewJournal of biophotonics2026

Two-Photon Fluorescence and Second-Harmonic Generation Microscopy: Principles and Biomedical Applications.

Kausalya Neelavara Makkithaya, Gagan Raju, Subir Das, Ajeetkumar Patil, Rajesh Kumar, Yury V Kistenev, Guan-Yu Zhuo, Nirmal Mazumder

Abstract readReview
In one paragraph

Review in Journal of biophotonics, 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.

0numbers the graph read from it
0cells of the map it votes in
0citing papers in PubMed
–field-weighted citation impact
1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

0 citing papers in PubMed.

No citing paper in PubMed yet.

4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

8 authors.

Kausalya Neelavara MakkithayaDepartment of Biophysics, Manipal School of Life Sciences, Manipal Academy of Higher Education, Manipal, India.
Gagan RajuDepartment of Biophysics, Manipal School of Life Sciences, Manipal Academy of Higher Education, Manipal, India.
Subir DasDepartment of Chemistry, University of Zurich, Zurich, Switzerland.
Ajeetkumar PatilManipal Institute of Applied Physics, Manipal Academy of Higher Education, Manipal, India.ORCID https://orcid.org/0000-0002-0790-9490
Rajesh KumarDepartment of Biomedical Engineering, Indian Institute of Technology, Ropar, India.ORCID https://orcid.org/0000-0001-8495-8838
Yury V KistenevLaboratory of Biophotonics, Tomsk State University, Tomsk, Russia.ORCID https://orcid.org/0000-0001-5760-1462
Guan-Yu ZhuoInstitute of Biophotonics, National Yang Ming Chiao Tung University, Taipei, Taiwan.ORCID https://orcid.org/0000-0002-9813-2989
Nirmal MazumderDepartment of Biophysics, Manipal School of Life Sciences, Manipal Academy of Higher Education, Manipal, India.ORCID https://orcid.org/0000-0001-8068-6484

Funding

Anusandhan National Research Foundation ANRF/F/84/2025-2026ASEAN-India Science & Technology Development Fund (AISTDF)Council of Scientific and Industrial Research, Government of India, India 08/0602(23331)/2025-EMR-IIndian Council of Medical Research D/SG-00170/2024
6 · The paper itself

Abstract

Two-photon microscopy (TPM) has revolutionized biological imaging by enabling high-resolution, deep-tissue visualization with reduced photobleaching and photodamage. This review discusses complementary nonlinear optical imaging modalities such as two-photon fluorescence (TPF), second-harmonic generation (SHG), and fluorescence lifetime imaging (FLIM) microscopy that together provide structural, molecular, and metabolic information. The review begins by introducing the basic principles, instrumentation, and capabilities of two-photon fluorescence microscopy and FLIM. Followed by the development of SHG microscopy, highlighting advances from conventional SHG imaging to polarization-resolved, circular dichroism, and Stokes-Mueller polarimetric approaches. The biomedical applications of these complementary techniques are subsequently discussed, followed by an overview of multimodal imaging strategies that combine structural, molecular, and functional information. Collectively, these modalities represent a versatile and rapidly evolving imaging toolkit with broad potential to advance biomedical research, disease diagnosis, and precision medicine.

Indexed as

Microscopy, Fluorescence, MultiphotonSecond Harmonic Generation MicroscopyAnimalsHumansbiomedical researchfluorescence lifetime imagingnonlinear optical microscopysecond‐harmonic generation microscopytwo‐photon microscopy

Identifiers

PMID42810836
PMCPMC13623537

What Socratic holds

Textmetadata
Read underepoch 390

Registered trials

None linked

Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the Socratic graph.