Why is it difficult to stain dormant bacteria with Nucleic acid dyes?

[littledog]

TL;DR

The correlation between bacterial physiological activity and Nucleic acid staining (SYTO) efficiency?

I found that it's difficult to stain dormant bacteria or bacteria in lag phase with Nucleic acid dye like SYTO9/SYBR Green Ⅰ, does anyone know why?
DNA 3D structure too complex? DNA binding protein too much? Low material transport efficiency in bacteria? Or anyother factors?
Is there any research about this?

 

[jim mcnamara]

This article agrees with your assessment of less staining for inactive bacteria.
https://academic.oup.com/femsec/article/29/4/319/526255
The authors mention membrane permeability.
@Ygggdrasil may have more information.

 

[littledog]

This article agrees with your assessment of less staining for inactive bacteria.
https://academic.oup.com/femsec/article/29/4/319/526255
The authors mention membrane permeability.
@Ygggdrasil may have more information.

I have try some methods to increase membrane permeability, the result shows that few effect on staining efficiency to inactive bacteria.

 

[Ygggdrasil]

Actively dividing bacteria are replicating their DNA and could have 2x or more DNA/cell than dormant bacteria that are not replicating.

 

[littledog]

Actively dividing bacteria are replicating their DNA and could have 2x or more DNA/cell than dormant bacteria that are not replicating.

But as what I observe in my experiment，there are a lot of bacteria in dividing phase couldn't be stained by SYTO13，and what I wonder is whether the DNA structure or I should say the density of DNA and its binding protein in dormant bacteria greatly effect the affinity between DNA and nucleic dye？

 

[BillTre]

Have you asked the dye manufacturer?
Do other DNA dyes have the same effect?

 

[littledog]

Have you asked the dye manufacturer?
Do other DNA dyes have the same effect?

YES，I almost buy all the DNA dyes of Thermo Fisher.