microscope calibration

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Henry Kuska

Re: microscope calibration

Post: # 13761Post Henry Kuska
Thu Oct 11, 2007 1:00 am

Looks like Amber Sun is a very infertile triploid. The first 2 links below are to what appears to be healthy 2N pollen grains and the last 2 links are to healthy 1N pollen grains, but most grains did not take up the stain.

Two 2N pollen

http://picasaweb.google.com/HAKuska/Mic ... 7026920466

http://picasaweb.google.com/HAKuska/Mic ... 9911822370

Two 1N pollen

http://picasaweb.google.com/HAKuska/Mic ... 8501756978

http://picasaweb.google.com/HAKuska/Mic ... 1386658882

Henry Kuska

Re: microscope calibration

Post: # 13768Post Henry Kuska
Fri Oct 12, 2007 1:00 am

David Zlesak has reported that Nearly Wild is a triploid. I have a (Nearly Wild OP) seedling. It is my number 293.

http://picasaweb.google.com/HAKuska/Hen ... 1687155666

From its pollen measurements, it appears to be a tetraploid.

http://picasaweb.google.com/HAKuska/Mic ... 1823108194

http://picasaweb.google.com/HAKuska/Mic ... 0413042802

Based on this information, I will have to make an attempt to cross it with some of my favorite tetraploids next season.

Link: picasaweb.google.com/HAKuska/HenrySRoses/photo#5076308151687155666


Timothy

Re: microscope calibration

Post: # 13786Post Timothy
Mon Oct 15, 2007 1:00 am

Thanks for the information!!

Thinking of buying my own microscoop.

I've seen one for 80 Euro. It's a basic one.

http://www.weberintershop.nl/images/pic ... /75925.jpg

40x, 100x en 400x

Henry Kuska

Re: microscope calibration

Post: # 13815Post Henry Kuska
Fri Oct 19, 2007 1:00 am

The following links are to pictures of Home Run pollen.

Http://picasaweb.google.com/HAKuska/Mic ... 9024295122

Http://picasaweb.google.com/HAKuska/Mic ... 3319262434

Http://picasaweb.google.com/HAKuska/Mic ... 7614229746

http://picasaweb.google.com/HAKuska/Mic ... 1909197058

Since both 1X and 2X pollen are present, I would say that Home Run is probably a triploid.

Robert Neil Rippetoe

Re: microscope calibration

Post: # 13816Post Robert Neil Rippetoe
Fri Oct 19, 2007 1:00 am

Thanks for this Henry. This explains why it's likely some of my Home Run seedlings are also triploid or appear to be. A few seem to be forming OP hips. There is hope. So far in general they look pretty good.

Any guess as to percentage of 1X opposed to 2X pollen?

Henry Kuska

Re: microscope calibration

Post: # 13862Post Henry Kuska
Mon Oct 22, 2007 1:00 am

Neil, I did not have a lot of pollen to work with as the flower had already opened.

The link below suggests that their would be sufficient 2X pollen to sucessfully pollinate a tetraploid.

http://picasaweb.google.com/HAKuska/Mic ... 2345921810

Link: picasaweb.google.com/HAKuska/MicroscopePictures02/photo#5124311972345921810

Robert Neil Rippetoe

Re: microscope calibration

Post: # 13868Post Robert Neil Rippetoe
Mon Oct 22, 2007 1:00 am

This is fascinating Henry. It makes me wish I had the patience to put something like this together. I think it will come in really handy, especially in estimating what to expect from the offspring of wide crosses.

Henry Kuska

Re: microscope calibration

Post: # 13883Post Henry Kuska
Tue Oct 23, 2007 1:00 am

I decided to see what a 2 megapixel webcam would do. Although 2 megapixel webcams are still very rare and expensive, the original Hewlett-Packard 2 Megapixel Webcam (with manual focus) has been replaced with one with automatic focus. Because of this model change, I was able to find the manual focus model for only $39.99, new.

Having a manual focus is a plus for microscope use as (in my experience) it has meant that the lens simply screws out.

The links below give comparison pictures of my Logitech Pro 4000 (a 640 X 480 CCD sensor) and the 2 Megapixel HP (a 1600 X 1200 CMOS sensor).

---------------------------------------------

HP 2-Megapixel webcam, calibration slide, 500X setting,1600 X 1200 pixels, 0.0640 microns per pixel Http://picasaweb.google.com/HAKuska/Mic ... 5310990626

---------------------------------------------

HP 2-Megapixel webcam, calibration slide, 100X setting,1600 X 1200 pixels, 0.285 microns per pixel

Http://picasaweb.google.com/HAKuska/Mic ... 3900925234

---------------------------------------------

Logitech Pro 4000 webcam, calibration slide, 500X setting,640 X 480 pixels, 0.108 microns per pixel

http://picasaweb.google.com/HAKuska/Mic ... 4612770146

---------------------------------------------

Logitech Pro 4000 webcam, calibration slide, 100X setting,640 X 480 pixels, 0.481 microns per pixel

Http://picasaweb.google.com/HAKuska/Mic ... 6022835538

----------------------------------------------

Timothy

Re: microscope calibration

Post: # 13987Post Timothy
Sat Nov 03, 2007 1:00 am

Is 300x enough to view cells and chromosomes?

David Zlesak

Re: microscope calibration

Post: # 13990Post David Zlesak
Sat Nov 03, 2007 1:00 am

Hi Timothy, Yes. I like 400x better, but 300x should do. At 100x (10x eyepiece, 10x objective multiplied together for 100x) it is nice to find groups of cells and sometimes possible areas where one can find cells in metaphase. It is difficult for me to detect visible chromosomes at 100x unless they are very very beautiful spread cells. Then I bump up the magnification and scan from there with the 40x objective. Pollen is much easier to see and 300x should be less of a challenge than looking for chromosomes.

Sincerely,

David

Timothy

Re: microscope calibration

Post: # 13991Post Timothy
Sat Nov 03, 2007 1:00 am

Thanks david! It's a big help. Now i'm sure I buy someting that I can use for my goal.

Timothy

Re: microscope calibration

Post: # 14034Post Timothy
Wed Nov 14, 2007 1:00 am

I got my scope yesterday! The eye ocular has a anoying sort of pencil looking thing in my view. It's from the side to the middle. I hope i can remove it.

Henry Kuska

Re: microscope calibration

Post: # 14035Post Henry Kuska
Wed Nov 14, 2007 1:00 am

Timothy, if you use a web cam, you remove the ocular so it does not make any difference (mine had one also).

Timothy

Re: microscope calibration

Post: # 14048Post Timothy
Thu Nov 15, 2007 1:00 am

I removed the ocular and the pinn is not in the ocular. It's in the tube. I'll mail the fabricator and ask how I can do it without damaging to much.

Timo

Re: microscope calibration

Post: # 14584Post Timo
Thu Jan 17, 2008 1:00 am

I tried to make some microscope-pictures with my new eos 400D but can get it sharp. This is the best one from the serie. It's mould from seeds I cleaned yesterday.

[img]members.chello.nl/t.drent4/mould.jpg[/img];

I'm going to look for a webcam like henry.

ann

Re: microscope calibration

Post: # 14589Post ann
Thu Jan 17, 2008 1:00 am

Timo,

Part of your problem is that your light source may produce parallel light at the center, but out at the edges, you're getting a distortion (the blue edge at upper center.)

If you had a monocular, I'd suggest moving the lens UNDER the stage of the microscope up closer and to do it while focusing on a single flat line (like a hair that could be flattened between slide and coverslip.) (It probably would need to be racked up.) Also, you might have the substage aperture too wide. There's an optimum width for each ocular.

I used to be good at getting images sharp for 40x and 100x oculars.

Feel free to ask questions. You may be trying to photograph something that's too thick for that set of lenses.

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