Re: DAPI on a 405 laser and 150x 1.45 objective

Posted by Goodhouse, Joseph G. on
URL: http://confocal-microscopy-list.275.s1.nabble.com/3D-volume-rendering-and-measurement-tp3647068p3693413.html

405 works perfectly fine with either Dapi or Hoechst

 

From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Renato Mortara
Sent: Tuesday, September 22, 2009 9:39 AM
To: [hidden email]
Subject: DAPI on a 405 laser and 150x 1.45 objective

 

Hello,

 

I wonder if I could get some feedback on how two doubts;

 

1) How well does the solid-state 405 laser performs with DAPI-labeled samples (Cells, tissues, etc.) with not-so-UV-transparent objectives ?

 

I currently run an old BioRad 1024UV and with the Coherent 622 Inova laser (351/363 lines) even with  not-so-UV-transparent objectives such as a 63x PlanApo and a 100X 1.4 PlanApo (both from Zeiss), by using DAPI at a relatively high concentration (20-50 uM) we get very nice signal.

 

2) Has anyone tried the 150x 1.45 NA Olympus objective (presumably designed for TIRF) with conventional/confocal fluorescence ? Is the small WD a problem ?

 

 

Any input will be greatly appreciated,

 

Best regards,

Renato

 

Renato A. Mortara

Parasitology Division

UNIFESP - Escola Paulista de Medicina

Rua Botucatu, 862, 6th floor

São Paulo, SP

04023-062

Brazil

Phone: [phone]

Fax:     [phone]

email: [hidden email]

home page: www.ecb.epm.br/~ramortara