Evaluating single-particle tracking in Lactococcus lactis by photo-activation localization microscopy (sptPALM) (CAT#: STEM-MIT-0399-LJX)

Introduction

Lactic acid bacteria (LAB) are frequently used in food fermentation and are invaluable for the taste and nutritional value of the fermentation end-product. To gain a better understanding of underlying biochemical and microbiological mechanisms and cell-to-cell variability in LABs, single-molecule techniques such as single-particle tracking photo-activation localization microscopy (sptPALM) hold great promises but are not yet employed due to the lack of detailed protocols and suitable assays. The service qualitatively tests various fluorescent proteins including variants that are photoactivatable and therefore suitable for sptPALM measurements in Lactococcus lactis, a key LAB for the dairy industry.

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Principle Applications Procedure Materials Notes Related Products

Principle

Principles of photoactivated localization microscopy: By fitting the two-dimensional Gaussian function to determine the centroid of microscope-formed light spots, a single fluorescent source (such as a fluorescent group) can be located with high precision. The accuracy of the calculation to determine the centroid depends only on the number of photons collected, and the resolution scale can be tens of nanometers or smaller. To achieve this accuracy, the density of the fluorescent molecules being tested is required to be low enough that the spots of the two fluorescent groups are unlikely to overlap.

Applications

Applied in many areas of the life sciences

Procedure

1. Sampling
2. Preparation of slices
3. Staining (Select according to the specific experimental situation)
4. Observation

Materials

• Sample Type:
Lactococcus lactis

Notes

Operate in strict accordance with the operating procedures, and shall not arbitrarily change the operating procedures

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