microscope calibration

I really am excited for you and the digital images you can take with your microscope and hopefully share your exciting finds with us.

I just put a drop of acetocarmine on the slide and some pollen and put the coverslip over it. To help get pollen on the drop I often take a toothpick moistened a bit from the drop, rub it in my pollen container and then transfer it that way.

You can get 30ml of acetocarmine for $3.95. That should last a long long time. http://www.neosci.com/catalog.asp?sid=91932079&showID=2415&content=cn_showitem Acetocarmine is just acetic acid (vinegar) with carmine boiled in it and strained out. Carmine is from insects. It will help make chromsomoes look darker if you use a little iron too. You can just use an iron needle to work with and it will darken the stain and chromosomes as you work. For pollen it just helps with contrast. Once can see aborted pollen grains more easily because they are pink and kind of empty.

Sincerely,

David

The link shows the pollen of Carefree Sunshine (plus some Ferrous gluconate since this batch was used for pollinating).

Is the presence of 2 sizes of pollen a strong indication that Carefree Sunshine is a triploid?

http://picasaweb.google.com/HAKuska/MicroscopePictures02/photo#5107571445226315490

Link: picasaweb.google.com/HAKuska/MicroscopePictures02/photo#5107571445226315490

Carefree Sunshine is tetraploid. A picture of a cell of Carefree Sunshine is included in the article I wrote on root tip squashes. The mean I got for the pollen was 42.7 microns with a very small standard deviation of 2.4.

I suspect those small possible pollen grains you are seeing may not stain very well and prove to look somewhat transparent with acetocarmine. They probably are aborted. Stain and color intensity are nice features of acetocarmine and other common pollen stains like analine blue.

Triploids tend to have a lot of aborted pollen and stainable pollen of quite variable size. Many dark, solid looking pollen grains of triploids do not germinate or germinate at very low rates perhaps due to aneuploidy. Leen Leus in her thesis and others have looked at pollen viability using pollen germination assays for triploids.

Sincerely,

David

The “Pocket Cam” arrived yesterday. The lens was removable, but the default resolution when in web cam mode is only 320 X 240. The higher resolutions are available through their software; but when I use the free Micam measurement program, I can only use the 320 X 240 mode. I assume that one would have to modify the drivers to get around this problem. I do not anticipate freeing up the time to attempt this.

I was too hasty in dismissing the AIPTEX “Pocket Cam”.

For some unknown reason the default resolution in web cam mode is now 640 X 480.

http://picasaweb.google.com/HAKuska/MicroscopePictures02/photo#5110122740794908386


Also, the still camera mode is useful since the camera comes with a 1.5 inch color LCD viewer. The pictures can be taken at resolutions of:

2048 X 1536

http://picasaweb.google.com/HAKuska/MicroscopePictures02/photo#5110122736499941074


1600 X 1200

http://picasaweb.google.com/HAKuska/MicroscopePictures02/photo#5110122727910006466


AND

1280 X 960

http://picasaweb.google.com/HAKuska/MicroscopePictures02/photo#5110122551816347314


The above can be compared with the RadioShack 640 X 480 picture given below:

http://picasaweb.google.com/HAKuska/MicroscopePictures02/photo#5110122745089875698


As can be seen from the pictures, the different resolutions for the AIPTEX does not change the field of view. The RadioShack web cam gives a smaller field of view (higher magnification) than the AIPTEX. This is not a problem when using the AIPTEX as the microscope is a zoom microscope with its highest setting at 500X.

The AIPTEX (in the web cam mode)can be used directly with the Micam measurement program. In the still camera mode the pictures can be saved and then loaded into the Micam program.

This is an acetocarmine stained microscope view of pollen of number 158 ((Bonavista OP)OP).

Any comments?

Link: picasaweb.google.com/HAKuska/MicroscopePictures02/photo#5110476920978005778

The following is a acetocarmine stained microscope view of pollen of number 350 Folksinger X Illusion.

I should mention that for both of these last 2 views, I stirred the pollen-acetocarmine drop on the slide with an iron nail as apparently the acetocarmine needs traces of iron to work correctly.

I seal the top and bottom slides together with a thin line of super glue.

Link: picasaweb.google.com/HAKuska/MicroscopePictures02/photo#5110503141753347890

The following is a acetocarmine stained microscope view of pollen of Viking Queen.

I have grown Viking Queen for many years and it has never set open pollinated hips. I do not remember if I previously tried using it as a pollen parent. Help-Me-Find does not list any offspring either way.

I am now trying to use it as a parent with alpha naphthalene acetamide (see: http://www.rosehybridizers.org/forum/message.php?topid=12511#13004 ).

Folksinger X Illusion is expected to be a tetraploid. The pollen of Viking Queen are very similar so it appears that there is a good chance that it is also tetraploid.

Bonavista self pollinated offspring are expected to be diploids. It appears that there may be some dead pollen (shrunken) among the larger “live” diploid pollen.

Link: picasaweb.google.com/HAKuska/MicroscopePictures02/photo#5110644961573461842

The following is a acetocarmine stained microscope view of pollen of Will Alderman.

Will Alderman in hybridizing (for me) behaved as if it was a triploid. I interpret the pollen size as suggesting that it may be just a diploid.



Link: picasaweb.google.com/HAKuska/MicroscopePictures02/photo#5110827175560994658

The linked web address is one acetocarmine stained microscope view of pollen of Simon Fraser. David Zlesak has determined that Simon Fraser is a triploid. Thus the larger pollen is probably a tetraploid and the smaller one is either a diploid or a shrunken dead pollen.

The link that has to be copied and pasted into the address bar contains another picture taken at the same time. It shows what are probably 2 healthy diploid pollen grains of Simon Fraser.

http://picasaweb.google.com/HAKuska/MicroscopePictures02/photo#5110960469871026050

Link: picasaweb.google.com/HAKuska/MicroscopePictures02/photo#5110953834146553714

Man, I need to finish college so I can afford this stuff :confused: I would love to be able to do this on my own in my own time. There are so many to be contrasted to given lineage.

Examples: Carefree Marvel, Rosarium Utersen, White Drift, Baby Faurax, Coral Drift, Escapade, the multiple version of Rosa californica, Golden Wings, some random china hybrids, etc.

In fact, I saty away from a lot of OGR’s because Im always afraid that their mix of ploidy combined with their once-blooming nature would be a total waste of time/money/energy to work with. But some of them are so cool! But something like this could make an OGR project less daunting.

Hi Jadae,

I learned that Baby Fraurax is dipoid and Golden Wings is tetraploid.

I just got my Modern Roses 12 and find it very nice overall. One disappointment that I have is that as far as I could tell looking through a few cultivars where ploidy was once mentioned that they omit ploidy now on cultivars. They do have ploidy listed for species, however. This doesn’t make sense to me, especially since species are populations of individuals and there could be variability in ploidy. With asexually propagated cultivars, however, there should be one stable ploidy level (of course there always seems to be a rare exception). I emailed the editors with ploidy level of the additional cultivars I confirmed months ago so it can be included. Oh well, maybe in Modern Roses 13 ploidy for cultivars can return. I do like the ease of looking up roses by marketing name. I was feeling disappointed that the variety name (first three letters of breeders last name plus whatever) was omitted. I was thinking at first that would be a huge problem in looking up rose information, especially patents since the marketing name is often left out and without it we would have a harder time finding the patent and maybe additional marketing names can be added to the same rose later. They do list the patent numbers. The only downside would be the registered roses within the past year or so that have patents applied for. It may be a challenge finding the patents later without the variety name. I suppose we can search the catalogs to see if it is listed and do extra homework to figure that out outside of Modern Rose 12 and get to it. For the most part in Modern Roses 12 all the old variety names have been replaced with trademark/marketing names even in the parentages. There are some oversights though and within Modern Roses 12 with old variety names in the parentage making it a challenge to correspond the variety names of parents with their trademark/marketing names.

I also miss not having a searchable CD with photos like Modern Roses 11 had.

I know it was a lot of work to get Modern Roses 12 together and appreciate all the efforts made.

What are others impressions about Modern Roses 12?

Sincerely,

David

I am linkimg to 3 pictures of acetocarmine stained microscope views of pollen of (Martin Frobisher OP) X Will Alderman

Http://picasaweb.google.com/HAKuska/MicroscopePictures02/photo#5111261182006255538

Http://picasaweb.google.com/HAKuska/MicroscopePictures02/photo#5111261177711288226

http://picasaweb.google.com/HAKuska/MicroscopePictures02/photo#5111261169121353618

In 2 of the pictures it looks like there is one pollen that could be tetraploid.

I won’t bother getting MR12. It doesn’t make sense to invest in a reference that can’t be updated constantly like an online database. I also think breeder code names are an important tool.

I’ve asked HelpMeFind to start including ploidy information which can be gleaned from older versions of Modern Roses and other references.

David, if you’d like to contact the owner of HelpMeFind I can put you in touch with him if you haven’t been in touch already.

Henry,

Thank you so much for posting the microscopic views of pollen grains. That old proverb," A picture is worth a thousand words" certainly holds true in this case. Seeing is finally understanding.

Thank you,

Jim

The following are what I obtained for a (Darlow’s Enigma OP) seedling’s pollen:

Http://picasaweb.google.com/HAKuska/MicroscopePictures02/photo#5111329618015150066

Http://picasaweb.google.com/HAKuska/MicroscopePictures02/photo#5111329609425215458

http://picasaweb.google.com/HAKuska/MicroscopePictures02/photo#5111329605130248146

Thanks David. That will help a lot. i opted not to use either GW or BF this year, bit I will next year since I will know what waters I may tread.

As I was going through Modern Roses 12 again last night I found another challenge. Without the variety names it was difficult to really know what the parents are sometimes of a more recent cultivar. There are some names that refer to multiple roses, such as ‘Amanda’. When a more recent rose listed ‘Amanda’ as a parent it was difficult to know which one was used. That happened periodically in Modern Roses 11 too, but it seems like that has become a larger problem in Modern Roses 12 because the variety names being used in teh parentages helped to clarify that point much of the time.

Sincerely,

David

Yeah, Ive had that issue with ‘Amanda’ too.

The following are what I obtained for Sunny Knockout’s pollen:

Http://picasaweb.google.com/HAKuska/MicroscopePictures02/photo#5111642132720508978

http://picasaweb.google.com/HAKuska/MicroscopePictures02/photo#5111642115540639778


The average diameter for 5 measured Sunny Knockout pollen grains are: 36.05, 38.7, 41.2, 38.4, and 35.7 microns. The average of these 5 averages is 38.0.


The average diameter for the Folksinger X Illusion pollen grains was 40.0.

The average diameter diameter for Viking Queen pollen grains was 38.5.

((Bonavista OP) OP) had 2 sizes of pollen, 26.6 and 36.25.

Simon Fraser had 2 sizes of pollen, 25.2 and 35.3.

Martin Frobisher X Will Alderman had 2 sizes of pollen, 27.8 and 33.05.

Will Alderman pollen averaged 28.8.

Darlow’s Enigma pollen averaged 30.5.


Please Note: The above numbers are just first looks. Many more measurements would have to be made to have scientific confidence in the supposed “differences”.