ArticleNature protocols2011
Characterization of bacterial spore germination using phase-contrast and fluorescence microscopy, Raman spectroscopy and optical tweezers.
Article in Nature protocols, 2011. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 50 papers.
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Who cites it
50 citing papers in PubMed.
- Multimodal High-Throughput Screening Raman Spectroscopy for Label-Free Single-Cell Characterization of Recombinant Protein Production in Baculovirus Expression Vector Systems.Analytical chemistry · 2026Article
- Nutrient-induced germination of Bacillus subtilis spores exhibiting shared metabolic profiles in both TSB and AGFK media.Metabolomics : Official journal of the Metabolomic Society · 2026Article
- Optical analysis of bacterial metabolic activity and spore germination via formazan formation.Analytical sciences : the international journal of the Japan Society for Analytical Chemistry · 2026Article
- An Integrated Workflow for the Purification and Fluorescence Imaging of Clostridioides difficile Spores Using Lanthanide Beta-Diketone Complexes.Methods in molecular biology (Clifton, N.J.) · 2026Article
- Revisiting bacterial spore germination in the presence of peptidoglycan fragments.Journal of bacteriology · 2025Article
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- Strategies for effective high pressure germination or inactivation ofApplied and environmental microbiology · 2024Article
- UV-Induced Spectral and Morphological Changes in Bacterial Spores for Inactivation Assessment.The journal of physical chemistry. B · 2024Article
- Construction of Graphene@Ag-MLF composite structure SERS platform and its differentiating performance for different foodborne bacterial spores.Mikrochimica acta · 2023Article
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- Label-free mid-infrared photothermal live-cell imaging beyond video rate.Light, science & applications · 2023Article
- Hypervirulent R20291 Clostridioides difficile spores show disinfection resilience to sodium hypochlorite despite structural changes.BMC microbiology · 2023Article
- Identification and characterization of new proteins crucial for bacterial spore resistance and germination.Frontiers in microbiology · 2023Article
- Optical tweezers across scales in cell biology.Trends in cell biology · 2022Review
- Ultrafast Widefield Mid-Infrared Photothermal Heterodyne Imaging.Analytical chemistry · 2022Article
- Visualization of SpoVAEa Protein Dynamics in Dormant Spores ofMicrobiology spectrum · 2022Article
- Discrimination of Stressed and Non-Stressed Food-Related Bacteria Using Raman-Microspectroscopy.Foods (Basel, Switzerland) · 2022Article
- Facilitated endospore detection forEngineering in life sciences · 2022Article
- Moderate High-Pressure Superdormancy inApplied and environmental microbiology · 2022Article
Corrections and comments
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Authors and funding
6 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
This protocol describes a method combining phase-contrast and fluorescence microscopy, Raman spectroscopy and optical tweezers to characterize the germination of single bacterial spores. The characterization consists of the following steps: (i) loading heat-activated dormant spores into a temperature-controlled microscope sample holder containing a germinant solution plus a nucleic acid stain; (ii) capturing a single spore with optical tweezers; (iii) simultaneously measuring phase-contrast images, Raman spectra and fluorescence images of the optically captured spore at 2- to 10-s intervals; and (iv) analyzing the acquired data for the loss of spore refractility, changes in spore-specific molecules (in particular, dipicolinic acid) and uptake of the nucleic acid stain. This information leads to precise correlations between various germination events, and takes 1-2 h to complete. The method can also be adapted to use multi-trap Raman spectroscopy or phase-contrast microscopy of spores adhered on a cover slip to simultaneously obtain germination parameters for multiple individual spores.
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Registered trials
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 OpenQuestion graph.