Off topic - Microinjection device

[Pedro Camello]

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Hi all,

 

This is a non confocal  question. I´m trying to apply repeated
microinjections of caffeine to cells using a microinjection device from
Eppendorf (Femtojet) while determining calcium in the microscope. Our
problem is that using microinjection pipettes (tip inner diameter < 0.5 mm)
the tip gets blocked by debris from the medium (even perfusing cells). If I
use broader pipettes (let´s say 10-30 micrometers wide) the caffeine of the
tip is released prematurely. I also have a picospritzer II, but I don´t have
even the manual .

 

I would appreciate if any body in the list with some experience with any of
these equipments contact me off-list (or on-list)

 

 

Thanks

[sprag]

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Pedro,

You can filter your solution using a micro filter 0.2 um before filling the injection needle. This should remove most of the debris from your solution and prevent blocking the needle.

Soren

On 08/03/2011 09:59, Pedro J Camello wrote:

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Hi all,



This is a non confocal  question. I´m trying to apply repeated
microinjections of caffeine to cells using a microinjection device from
Eppendorf (Femtojet) while determining calcium in the microscope. Our
problem is that using microinjection pipettes (tip inner diameter < 0.5 mm)
the tip gets blocked by debris from the medium (even perfusing cells). If I
use broader pipettes (let´s say 10-30 micrometers wide) the caffeine of the
tip is released prematurely. I also have a picospritzer II, but I don´t have
even the manual .



I would appreciate if any body in the list with some experience with any of
these equipments contact me off-list (or on-list)





Thanks


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[Tim Feinstein-2]

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Hi Pedro,

One common practice is to spin solutions for, e.g., ~10 min. at 13k on a benchtop centrifuge just before loading the injection needle.  People often do this when small solution volume makes  filtering impractical. The last time I checked Eppendorf had a number of protocols available on their website that describe this.  

cheers,


TF

Timothy Feinstein, PhD
Postdoctoral Fellow
Laboratory for GPCR Biology
Dept. of Pharmacology & Chemical Biology
University of Pittsburgh, School of Medicine
BST W1301, 200 Lothrop St.
Pittsburgh, PA  15261

On Mar 8, 2011, at 4:59 AM, Pedro J Camello wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Hi all,
>
>
>
> This is a non confocal  question. I´m trying to apply repeated
> microinjections of caffeine to cells using a microinjection device from
> Eppendorf (Femtojet) while determining calcium in the microscope. Our
> problem is that using microinjection pipettes (tip inner diameter < 0.5 mm)
> the tip gets blocked by debris from the medium (even perfusing cells). If I
> use broader pipettes (let´s say 10-30 micrometers wide) the caffeine of the
> tip is released prematurely. I also have a picospritzer II, but I don´t have
> even the manual .
>
>
>
> I would appreciate if any body in the list with some experience with any of
> these equipments contact me off-list (or on-list)
>
>
>
>
>
> Thanks

[Carol Heckman]

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Pedro-
Are you using the femto-tips?  This are much, much smaller in inner diameter than 0.5 mm.  They can be purchased from Eppendorf or Zeiss.  I have used them but I cannot remember which was the vendor.
Center for Microscopy & Microanalysis
Bowling Green State University
________________________________________
From: Confocal Microscopy List [[hidden email]] On Behalf Of Pedro J Camello [[hidden email]]
Sent: Tuesday, March 08, 2011 4:59 AM
To: [hidden email]
Subject: Off topic - Microinjection  device

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Hi all,



This is a non confocal  question. I´m trying to apply repeated
microinjections of caffeine to cells using a microinjection device from
Eppendorf (Femtojet) while determining calcium in the microscope. Our
problem is that using microinjection pipettes (tip inner diameter < 0.5 mm)
the tip gets blocked by debris from the medium (even perfusing cells). If I
use broader pipettes (let´s say 10-30 micrometers wide) the caffeine of the
tip is released prematurely. I also have a picospritzer II, but I don´t have
even the manual .



I would appreciate if any body in the list with some experience with any of
these equipments contact me off-list (or on-list)





Thanks