Re: Syto dyes for plant nuclei in living cells VENDOR RESPONSE

[Roy Edward]

Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

Hi John

There was a paper in The Plant Cell (Da Costa et al, The Plant Cell 2006; 18:1626-1437) that describes use of DRAQ5 for Arabidopsis root hair nuclei:

http://www.plantcell.org/cgi/content/abstract/18/6/1426

The image for this can be found in the paper's supplementary information:

http://www.plantcell.org/cgi/data/tpc.105.040485/DC1/2

Regards

Roy

 

Roy Edward

Biostatus Ltd

TEL: [phone]; [hidden email]; www.biostatus.com

Company No: 3079239. Regd. in England & Wales.

Regd. office: 56 Charnwood Road, Shepshed, LE12 9NP, UK.

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From: John Runions [mailto:[hidden email]]
Sent: Thu 08/05/2008 11:13
Subject: Syto dyes for plant nuclei in living cells

Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

Do any of the plant biologists out there have experience using the Syto dyes from Molecular Probes to stain nuclei of living cells.  In particular, they list 5 different dyes in the blue emission range (Syto 40-45).  Syto 42 looks good to me on paper.  What do you think.  Thanks, John.

[John Runions]

Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

Thanks Roy, the images in DaCosta et al make DRAQ5 staining of living plant nuclei look pretty convincing.  I can even see dividing cells!  They don't provide much info about its penetration into the cell layers of the root.  Has anybody else tried DRAQ5?  John.

Roy Edward wrote:

Hi John

There was a paper in The Plant Cell (Da Costa et al, The Plant Cell 2006; 18:1626-1437) that describes use of DRAQ5 for Arabidopsis root hair nuclei:

http://www.plantcell.org/cgi/content/abstract/18/6/1426

The image for this can be found in the paper's supplementary information:

http://www.plantcell.org/cgi/data/tpc.105.040485/DC1/2

Regards

Roy

 

Roy Edward

Biostatus Ltd

TEL: [phone]; [hidden email]; www.biostatus.com

Company No: 3079239. Regd. in England & Wales.

Regd. office: 56 Charnwood Road, Shepshed, LE12 9NP, UK.

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From: John Runions [[hidden email]]
Sent: Thu 08/05/2008 11:13
Subject: Syto dyes for plant nuclei in living cells

Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

Do any of the plant biologists out there have experience using the Syto dyes from Molecular Probes to stain nuclei of living cells.  In particular, they list 5 different dyes in the blue emission range (Syto 40-45).  Syto 42 looks good to me on paper.  What do you think.  Thanks, John.

[Tobias Baskin]

Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

John,

        Yes, we also tried DRAQ5 (Roy Edward was very helpful). But were not able to get useful results (we needed to label cells in the root body rather than root hair). We did contact Da Costa et al. to find out about any "secrets" but they weren't aware of any. Must be the water in Belgium or something like that? Sorry to spread more rain.

        Tobias

Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

Thanks Roy, the images in DaCosta et al make DRAQ5 staining of living plant nuclei look pretty convincing.  I can even see dividing cells!  They don't provide much info about its penetration into the cell layers of the root.  Has anybody else tried DRAQ5?  John.

Roy Edward wrote:

Runions signature

Hi John

There was a paper in The Plant Cell (Da Costa et al, The Plant Cell 2006; 18:1626-1437) that describes use of DRAQ5 for Arabidopsis root hair nuclei:

http://www.plantcell.org/cgi/content/abstract/18/6/1426

The image for this can be found in the paper's supplementary information:

http://www.plantcell.org/cgi/data/tpc.105.040485/DC1/2

Regards

Roy

 

Roy Edward

Biostatus Ltd

TEL: [phone]; [hidden email]; www.biostatus.com

Company No: 3079239. Regd. in England & Wales.

Regd. office: 56 Charnwood Road, Shepshed, LE12 9NP, UK.

---

From: John Runions [[hidden email]]
Sent: Thu 08/05/2008 11:13
Subject: Syto dyes for plant nuclei in living cells

Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

Do any of the plant biologists out there have experience using the Syto dyes from Molecular Probes to stain nuclei of living cells.  In particular, they list 5 different dyes in the blue emission range (Syto 40-45).  Syto 42 looks good to me on paper.  What do you think.  Thanks, John.

--

*********************************
C. John Runions, Ph.D.
School of Life Sciences
Oxford Brookes University
Oxford, UK
OX3 0BP

email:  [hidden email]
phone: [phone]

Runions' lab web site

 

Visit The Illuminated Plant Cell dot com
Oxford Brookes Master's in Bioimaging with Molecular Technology

--
Runions signature

*********************************
C. John Runions, Ph.D.
School of Life Sciences
Oxford Brookes University
Oxford, UK
OX3 0BP

email:  [hidden email]
phone: [phone]

Runions' lab web site

 

Visit The Illuminated Plant Cell dot com
Oxford Brookes Master's in Bioimaging with Molecular Technology

-- 

      _      ____          __   ____   

     /  \   /          / \    /   \ \        Tobias I. Baskin

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[Roy Edward]

Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

Hi John

This is the first paper I have seen on plant tissue per se.

There was a paper in Planta by Libault et al (2005) that did some work on a GFP transgenic Nicotiana cell line for mitosis studies. 

We have done some work ourselves on fungal hyphae that looked very similar to these root hairs!!

I guess, though, that the big issue is the cell wall with respect to penetration of layers. 

DRAQ5 appears to penetrate quite deep into mammalian tissue layers/spheroids and is more readily detectable since it's excited in the red at 633 or 647nm.

 

If this is still interesting and worth pursuing by your lab the ordering info is on our website.  If despite our tech support efforts, it doesn't work we will refund the cost of the DRAQ5. If it does work then let's see if there's a methods poster to present somewhere!!

 

Best regards

Roy

 

Roy Edward

Sales & Mktg. Director, Biostatus Ltd

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From: John Runions [mailto:[hidden email]]
Sent: Fri 09/05/2008 11:00
To: Roy Edward; Confocal Microscopy List
Subject: DRAQ5 for plant nuclei in living cells

Thanks Roy, the images in DaCosta et al make DRAQ5 staining of living plant nuclei look pretty convincing.  I can even see dividing cells!  They don't provide much info about its penetration into the cell layers of the root.  Has anybody else tried DRAQ5?  John.

Roy Edward wrote:

Hi John

There was a paper in The Plant Cell (Da Costa et al, The Plant Cell 2006; 18:1626-1437) that describes use of DRAQ5 for Arabidopsis root hair nuclei:

http://www.plantcell.org/cgi/content/abstract/18/6/1426

The image for this can be found in the paper's supplementary information:

http://www.plantcell.org/cgi/data/tpc.105.040485/DC1/2

Regards

Roy

 

Roy Edward

Biostatus Ltd

TEL: [phone]; [hidden email]; www.biostatus.com

Company No: 3079239. Regd. in England & Wales.

Regd. office: 56 Charnwood Road, Shepshed, LE12 9NP, UK.

---

From: John Runions [[hidden email]]
Sent: Thu 08/05/2008 11:13
Subject: Syto dyes for plant nuclei in living cells

Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

Do any of the plant biologists out there have experience using the Syto dyes from Molecular Probes to stain nuclei of living cells.  In particular, they list 5 different dyes in the blue emission range (Syto 40-45).  Syto 42 looks good to me on paper.  What do you think.  Thanks, John.

[Rosemary.White]

Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal Hi Roy, John,
Yes, I’ve tried DRAQ5, it gets into external cells fine, but doesn’t penetrate very deeply very quickly.  It gets into some cells but not others, like DAPI, but is handy to have a red emitting nuclear stain.  Like any other DNA stain I’ve tried, it also labels cell walls.  An additional feature is that strongly stained nuclei appear blue in brightfield, my guess is that more strongly stained cells, e.g. sloughing root cap cells, root hair cells, have more permeable membranes or their membranes have been compromised in their trip from growth medium to microscope slide.

cheers,
Rosemary

On 9/5/08 7:50 PM, "John Runions" <[hidden email]> wrote:

Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

Thanks Roy, the images in DaCosta et al make DRAQ5 staining of living plant nuclei look pretty convincing.  I can even see dividing cells! They don't provide much info about its penetration into the cell layers of the root.  Has anybody else tried DRAQ5?  John.

Roy Edward wrote:

Runions signature   
 
Hi John
 
There was a paper in The Plant Cell (Da Costa et al, The Plant Cell 2006; 18:1626-1437) that describes use of DRAQ5 for Arabidopsis root hair nuclei:
 
http://www.plantcell.org/cgi/content/abstract/18/6/1426
 
The image for this can be found in the paper's supplementary information:
 
http://www.plantcell.org/cgi/data/tpc.105.040485/DC1/2
 
Regards
 
Roy
 
 
 
 
 
 
Roy Edward

Biostatus Ltd

TEL: [phone]; [hidden email] [hidden email] ; www.biostatus.com <http://www.biostatus.com/>
 
Company No: 3079239. Regd. in England & Wales.

Regd. office: 56 Charnwood Road, Shepshed, LE12 9NP, UK.

 
 

 

---

From: John Runions [[hidden email]
 Sent: Thu 08/05/2008 11:13
 Subject: Syto dyes for plant nuclei in living cells
 

 
Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
 
Do any of the plant biologists out there have experience using the Syto dyes from Molecular Probes to stain nuclei of living cells.  In particular, they list 5 different dyes in the blue emission range (Syto 40-45).  Syto 42 looks good to me on paper.  What do you think. Thanks, John.
 

[Scott Snyder]

Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal I used to use DRAQ5 a lot.  I agree that staining can be variable but I've seen variation in apparently healthy cultured cells.  I have not figured out the pattern to it as yet but when the people who make it do, I think they will have a lot more success because there is definitely a specificity to it.  It will also interact directly with BODIPY containing dyes so be careful there.

Scott Snyder

Rosemary White <[hidden email]> wrote:

Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal Hi Roy, John,
Yes, I’ve tried DRAQ5, it gets into external cells fine, but doesn’t penetrate very deeply very quickly.  It gets into some cells but not others, like DAPI, but is handy to have a red emitting nuclear stain.  Like any other DNA stain I’ve tried, it also labels cell walls.  An additional feature is that strongly stained nuclei appear blue in brightfield, my guess is that more strongly stained cells, e.g. sloughing root cap cells, root hair cells, have more permeable membranes or their membranes have been compromised in their trip from growth medium to microscope slide.

cheers,
Rosemary

On 9/5/08 7:50 PM, "John Runions" <[hidden email]> wrote:

Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

Thanks Roy, the images in DaCosta et al make DRAQ5 staining of living plant nuclei look pretty convincing.  I can even see dividing cells! They don't provide much info about its penetration into the cell layers of the root.  Has anybody else tried DRAQ5?  John.

Roy Edward wrote:

Runions signature   
 
Hi John
 
There was a paper in The Plant Cell (Da Costa et al, The Plant Cell 2006; 18:1626-1437) that describes use of DRAQ5 for Arabidopsis root hair nuclei:
 
http://www.plantcell.org/cgi/content/abstract/18/6/1426
 
The image for this can be found in the paper's supplementary information:
 
http://www.plantcell.org/cgi/data/tpc.105.040485/DC1/2
 
Regards
 
Roy
 
 
 
 
 
 
Roy Edward

Biostatus Ltd

TEL: [phone]; [hidden email] [hidden email] ; www.biostatus.com <http://www.biostatus.com/>
 
Company No: 3079239. Regd. in England & Wales.

Regd. office: 56 Charnwood Road, Shepshed, LE12 9NP, UK.

 
 

 

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From: John Runions [[hidden email]
 Sent: Thu 08/05/2008 11:13
 Subject: Syto dyes for plant nuclei in living cells
 

 
Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
 
Do any of the plant biologists out there have experience using the Syto dyes from Molecular Probes to stain nuclei of living cells.  In particular, they list 5 different dyes in the blue emission range (Syto 40-45).  Syto 42 looks good to me on paper.  What do you think. Thanks, John.