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Thank you everyone for all the advice and comments in regards to my  
original question. The references were very helpful too. Looks like  
I'll have to go back to TIRF.

John

On 31-Jul-07, at 10:29 AM, Michelle Peckham wrote:

> Search the CONFOCAL archive at
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
> Or see:
>
> Visualizing single molecules inside living cells using total internal
> reflection fluorescence microscopy.
>
> Mashanov GI, Tacon D, Knight AE, Peckham M, Molloy JE. (2003)
>
> Methods  Feb;29(2):142-52.
>
> And
>
> Automatic detection of single fluorophores in live cells.
>
> Mashanov GI, Molloy JE.
>
> Biophys. J. (2007) 92(6) 2199-211
>
> Gregory has developed nice algorithms for detecting stepwise  
> photobleaching,
> with the aim of detecting signle fluorphores,  which he is happy  
> for anyone
> to use, reference is in the second of the papers.
>
> Michelle
>
>
> On 31/7/07 13:55, "Ian Dobbie" <[log in to unmask]> wrote:
>
>> Search the CONFOCAL archive at
>> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>>
>> David Knecht-charter <[log in to unmask]> writes:
>>
>>> Search the CONFOCAL archive at
>>> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>>>
>>> Given Martin's comment, what is a good simple and conclusive test  
>>> for
>>> determining sensitivity in the 1-10 fluorophore/molecule range if
>>> testing cameras/systems?  Dave
>>
>> In a TIRF system you should be able to get stepwise photobleaching to
>> see how many fluorophores you have:
>> Nature 443, 355-358 (2006)
>> Stoichiometry and turnover in single, functioning membrane protein  
>> complexes
>> Mark C. Leake, Jennifer H. Chandler, George H. Wadhams, Fan Bai1,  
>> Richard M.
>> Berry  and  Judith P. Armitage
>>
>>
>> Ian