Title of the invention: A method for live cell imaging of bacterial cells using anionic porphyrins and its metallo derivatives
Date of filing : 11/05/2009
Application No : 1082/CHE/2009
Name of the applicant : P. Gautam, V. Sujatha, K.S.Vinod Kumar, SrinathKrishnamurthy and Bharat Sridhar
Name of inventor : P. Gautam, V. Sujatha, K.S.Vinod Kumar, SrinathKrishnamurthy and Bharat Sridhar
The main objective of the present invention is live cell imaging
technique for bacterial cells using anionic porphyrins and its metallo
derivatives. In a specific illustrative preferred embodiment of this aspect of the
invention, Pseudomonas culture was grown for 24hrs at 37 ° C at 160 rpm in
Luria-Bertani medium using standard microbiological procedure.
One ml of the Pseudomonas culture was mixed with \Q\x] of anionic
porphyrin and 10 \x\ of zinc, derivative in separate eppendorf tubes under
aseptic condition. The sample was incubated in room temperature for 5 min.
One drop of the sample was taken in a glass slide and was visualized under
ii TIRF microscope, where the organisms fluoresced well. The fluorescence of
the moving cells indicates that the organism has taken up the fluorophore.
The non-toxic nature of the fluorophore is quite promising in extending
this procedure for fluorescent studying localization, trafficking, and expression
levels of bio molecules in live cells.