# Advanced Imaging for Live-Cell Spatiotemporal Monitoring: Technologies and Applications

**Authors:** Zhong Zhuang, Zhichao Feng, Jie Wang, Xinhui Liu, Laijun Song, Chunhui Sun, Hong Liu, Na Ren

PMC · DOI: 10.34133/research.1085 · Research · 2026-02-10

## TL;DR

This review discusses advanced imaging technologies that allow high-resolution, real-time observation of live-cell processes, offering new insights into cellular behavior and potential applications in medicine.

## Contribution

The paper provides a comprehensive overview of four cutting-edge imaging techniques and their roles in live-cell spatiotemporal monitoring.

## Key findings

- Advanced imaging technologies enable precise tracking of molecular interactions and cellular dynamics at the nanoscale.
- These methods have significantly improved the ability to observe live-cell processes like division and signaling.
- Integration of these tools into biomedical research promises better disease diagnosis and drug discovery.

## Abstract

Understanding cellular dynamics requires real-time high-resolution imaging. Recent advancements in imaging technologies have provided unprecedented spatial and temporal resolutions, enabling the precise in situ monitoring of live-cell behavior. This review covers 4 advanced imaging modalities: stimulated emission depletion microscopy, structured illumination microscopy, single-molecule localization microscopy, and Raman spectroscopy. We summarize the principles, applications, advantages, and limitations of these methods, highlighting their significance for high-precision spatiotemporal monitoring of cellular structures and biochemical activities. These tools enable precise tracking of molecular interactions and analysis of cellular dynamics at the nanoscale, which is critical for understanding cellular physiology. The integration of these technologies into biomedical research has markedly enhanced our ability to observe live-cell processes, such as division, migration, differentiation, and signaling. The development and application of these high-precision imaging technologies hold substantial promise for improving disease diagnosis, therapeutic strategies, and drug discovery.

## Full-text entities

- **Genes:** SPINK5 (serine peptidase inhibitor Kazal type 5) [NCBI Gene 11005] {aka LEKTI, LETKI, NETS, NS, VAKTI}, LAMP1 (lysosome associated membrane protein 1) [NCBI Gene 3916] {aka CD107a, LAMPA, LGP120}, App (amyloid beta precursor protein) [NCBI Gene 11820] {aka Abeta, Abpp, Adap, Ag, Cvap, E030013M08Rik}, Selp (selectin, platelet) [NCBI Gene 20344] {aka CD62P, GMP-140, Grmp, LECAM3, PADGEM}, EEA1 (early endosome antigen 1) [NCBI Gene 8411] {aka MST105, MSTP105, ZFYVE2}, VIM (vimentin) [NCBI Gene 7431], PALM (paralemmin) [NCBI Gene 5064] {aka PALM1}, EREG (epiregulin) [NCBI Gene 2069] {aka EPR, ER, Ep}, APP (amyloid beta precursor protein) [NCBI Gene 351] {aka AAA, ABETA, ABPP, AD1, APPI, CTFgamma}, CD63 (CD63 molecule) [NCBI Gene 967] {aka AD1, HOP-26, ME491, MLA1, OMA81H, Pltgp40}, XIST (X inactive specific transcript) [NCBI Gene 7503] {aka DXS1089, DXS399E, LINC00001, NCRNA00001, SXI1, swd66}, FADS2 (fatty acid desaturase 2) [NCBI Gene 9415] {aka D6D, DES6, FADSD6, LLCDL2, SLL0262, TU13}, EGFR (epidermal growth factor receptor) [NCBI Gene 1956] {aka ERBB, ERBB1, ERRP, HER1, NISBD2, NNCIS}, H2BC21 (H2B clustered histone 21) [NCBI Gene 8349] {aka GL105, H2B, H2B-GL105, H2B.1, H2BE, H2BFQ}, CYP3A4 (cytochrome P450 family 3 subfamily A member 4) [NCBI Gene 1576] {aka CP33, CP34, CYP3A, CYP3A3, CYPIIIA3, CYPIIIA4}, SCD (stearoyl-CoA desaturase) [NCBI Gene 6319] {aka FADS5, MSTP008, SCD1, SCDOS, hSCD1}, PEX11A (peroxisomal biogenesis factor 11 alpha) [NCBI Gene 8800] {aka PEX11-ALPHA, PEX11alpha, PMP28, hsPEX11p}, SLC6A3 (solute carrier family 6 member 3) [NCBI Gene 6531] {aka DAT, DAT1, PKDYS, PKDYS1}, IL6 (interleukin 6) [NCBI Gene 3569] {aka BSF-2, BSF2, CDF, HGF, HSF, IFN-beta-2}, RYR2 (ryanodine receptor 2) [NCBI Gene 6262] {aka ARVC2, ARVD2, RYR-2, RyR, VACRDS, VTSIP}, TUBA1B (tubulin alpha 1b) [NCBI Gene 10376] {aka K-ALPHA-1}
- **Diseases:** glycogen metabolism disorder (MESH:D008659), SMLM (MESH:D012640), gastric cancer (MESH:D013274), phototoxic (MESH:D017484), neurological diseases (MESH:D020271), cancer (MESH:D009369), AD (MESH:D000544), lung cancer (MESH:D008175), STORM (MESH:D009901), prostate cancer (MESH:D011471), neurodegeneration (MESH:D019636), mitochondrial diseases (MESH:D028361), esophageal cancer (MESH:D004938), TC (OMIM:275350), glomerular diseases (MESH:D007674), breast cancer (MESH:D001943), SR-SMLM (MESH:C535318), cytotoxicity (MESH:D064420), hallucinations (MESH:D006212), colorectal cancer (MESH:D015179), EC (MESH:D005955), thyroid cancer (MESH:D013964)
- **Chemicals:** adenine (MESH:D000225), glutamate (MESH:D018698), SC 26196 (MESH:C120986), PDA (MESH:C568283), Ag (MESH:D012834), galactose (MESH:D005690), cholesterol (MESH:D002784), SC (MESH:D012538), cardiolipin (MESH:D002308), neodymium (MESH:D009354), TMR (MESH:C005358), phospholipids (MESH:D010743), glycogen (MESH:D006003), PEG (MESH:D011092), EDTA (MESH:D004492), nitrogen (MESH:D009584), CO (MESH:D002248), carbon (MESH:D002244), polymer (MESH:D011108), ClO- (MESH:D006997), Au (MESH:D006046), metal (MESH:D008670), phenylazide (MESH:C014747), ZnSe (MESH:C044696), rhodamine (MESH:D012235), Nile Red (MESH:C044808), CAY10566 (MESH:C531040), oleate (MESH:D019301), lanthanide (MESH:D028581), Cy5 (MESH:C085321), ROS (MESH:D017382), calcium (MESH:D002118), sphingosine (MESH:D013110), Mn (MESH:D008345), glucose (MESH:D005947), 4',6-diamidino-2-phenylindole (MESH:C007293), OS (MESH:D009992), FM4-64 (MESH:C092350), Lipids (MESH:D008055), l-cysteine (MESH:D003545), cetyltrimethylammonium bromide (MESH:D000077286), CdTe (MESH:C028337), thiol (MESH:D013438), fatty acid (MESH:D005227), polyacetylene (MESH:D000078789), Phe (MESH:D010649), NO (MESH:D009614), H2S (MESH:D006862), deuterium (MESH:D003903), Alexa Fluor 532 (-), AF647 (MESH:C569686), sodium citrate (MESH:D000077559), silica (MESH:D012822)
- **Species:** Mus musculus (house mouse, species) [taxon 10090], Caenorhabditis elegans (species) [taxon 6239], Cricetus cricetus (black-bellied hamster, species) [taxon 10034], Staphylococcus aureus (species) [taxon 1280], Homo sapiens (human, species) [taxon 9606]
- **Mutations:** M159T
- **Cell lines:** VA-13 — Homo sapiens (Human), Hepatoblastoma, Cancer cell line (CVCL_A5CQ), PtK2 — Potorous tridactylus (Potoroo), Spontaneously immortalized cell line (CVCL_0514), HEK — Homo sapiens (Human), Human papillomavirus-related endocervical adenocarcinoma, Cancer cell line (CVCL_M624), H9c2 — Rattus norvegicus (Rat), Spontaneously immortalized cell line (CVCL_0286), M381 — Homo sapiens (Human), Melanoma, Cancer cell line (CVCL_XJ98), U2OS — Homo sapiens (Human), Osteosarcoma, Cancer cell line (CVCL_0042), 3T3-L1 — Mus musculus (Mouse), Spontaneously immortalized cell line (CVCL_0123), HepaRG — Homo sapiens (Human), Hepatitis C infection, Cancer cell line (CVCL_9720), Vero — Chlorocebus sabaeus (Green monkey), Spontaneously immortalized cell line (CVCL_0059), HeLa — Homo sapiens (Human), Human papillomavirus-related endocervical adenocarcinoma, Cancer cell line (CVCL_0030), U87 — Homo sapiens (Human), Glioblastoma, Cancer cell line (CVCL_0022), HEK293 — Homo sapiens (Human), Transformed cell line (CVCL_0045), HuH7 — Homo sapiens (Human), Adult hepatocellular carcinoma, Cancer cell line (CVCL_0336), MCF-10A — Homo sapiens (Human), Spontaneously immortalized cell line (CVCL_0598), BHK-21 — Mesocricetus auratus (Golden hamster), Spontaneously immortalized cell line (CVCL_RQ70), COS-7 — Chlorocebus aethiops (Green monkey), Transformed cell line (CVCL_0224), WI-38 — Homo sapiens (Human), Finite cell line (CVCL_0579)

## Full text

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## Figures

20 figures with captions in the complete paper: https://tomesphere.com/paper/PMC12886719/full.md

## References

200 references — full list in the complete paper: https://tomesphere.com/paper/PMC12886719/full.md

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Source: https://tomesphere.com/paper/PMC12886719