Re: Sorry-one more question...and a rant

Janet,

As someone once said, "I feel your pain! "   And how about the people who 
bring you huge experiments meant for publication without the controls that 
are necessary to set up the instrument and want you to "just make it look 
right"?  [ Actually, this might apply to your question about the cells that 
only fluoresce when viewed under a microscope, not when they are run 
through the flow cytometer. ]

Another question that I would ask if someone claimed that their "negative 
by flow" samples were "positive by microscopy" would be what filter blocks 
they were viewing their cells through, and what illumination source they 
used.  It may well be that they saw florescence, but it might not be the 
florescence of what they thought they were seeing.  Lots of things 
fluoresce if excited by the right wave length(s) and viewed though the 
right filters.

Of course, you could also offer to take a look at their sample under the 
microscope.  Seeing what they are talking about may give the necessary insight.

Hang in there.

Rick Meister


At 10:15 AM 7/15/04 -0400, you wrote:
>Dear Flowers.
>
>
>I now have another question and a little rant to all who I know will 
>appreciate it.     I have had several clients bring me samples which they 
>claim to have examined under fluorescence scope and can see their labeled 
>cells shining brightly.  We then run the sample through the machine and 
>can see little or no signal.
>
>Can someone give me a terribly scientific explanation for this that I can 
>rattle off to these people who want to blame me when their experiment 
>doesn't work???
>
>Okay here is the rant:
>
>
>Also is anyone else tired of being blamed when experiments don't 
>work?!?  I get really tired of having to try to help trouble shoot 
>experiments for people only to find out 20 minutes into the discussion 
>that this experiment has never been tried before or they are using an 
>antibody developed for a different species or they are using a 
>transvection vector developed for bacteria on yeast cells. They just can't 
>understand why it didn't work and it must be something that I am 
>doing.   But my favorite is when PIs send graduate students or post docs 
>to do all the work, and then contact me later because the experiments 
>haven't  worked and are mad because they are being charged for it.   Of 
>course, it is all my fault the experiments didn't work and how can I have 
>the nerve to charge them for it!!!
>
>
>Okay that is enough and I know I am preaching to the choir, so thanks for 
>listening.    The truth is I am really luck and my boss totally backs me 
>up and defends me relentlessly to these people.  The really fun part is my 
>coworkers in my other lab always know when I have just dealt with one of 
>these people as I will immediately go downstairs and get chocolate from 
>the special super candy bar fund in our basement.
>Thank god for chocolate and peanut butter.
>
>Again thanks for listening and for being the only ones who can truly 
>appreciate.
>
>Thanks again for all the replies
>
>Janet Dow
>
>
>--
>Janet Dow
>Laboratory Research Specialist and Manager
>Flow Cytometry and Cell Sorting Facility
>North Carolina State College of Veterinary Medicine
>Room C-309
>Raleigh, NC 27606
>(919)513-6443

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