🔬 063 - What PHASE CONTRAST microscopy is - all you ever wanted to know

[@MemoriesRR]

By far the most educational channel on KZbin when it comes to the microscope.. You use terms I can under stand and when a big word is used you explain it. You're not trying to impress but rather educate. Thank you so very much sir.

[@Microbehunter]

Thank you!

[@FL_Guns_N_Games]

Couldn’t agree more, I bought myself a decent beginners microscope because of this guy

[@footy7productions665]

Saaaame

[@DrFarazHarsini]

By far one of the best resources on KZbin when it comes to microscope. You get to the point, are excited, and give so much important information without rambling! Thanks a million for that!

[@paroutdiok_awesome]

I've been watching your videos for some days now and i was surprised to find out your views are not equivalent to the quality you put out. I hope more people find out about you!

[@christopherjob5332]

I use PCM for work as a CSST and this video really helped me understand a larger scope of application for my field equipment. Thank you!

[@suvidani]

You see the benefit of phase contrast when you are expecting to see some ultrastructure within the cells. Unfortunately the samples you have tried were all plant cross-sections which were not too rich in organelles in the first place, but the areas were crowded in cells, so at the end the field got messy without any obvious benefit. I think if you looked at unicellular protozoans or algae where the large cells are crowded with different kinds of vesicles the improvement would be stunning.

[@johnharvey4496]

Yes, get some pond water under the 'scope, they we'll all be sportin' it.

[@donaldeisenbarth5255]

I am very impressed with your channel. You have an excellent voice. Being older and with some hearing loss at certain frequencies I have difficulty understanding many channels. You speak so distinctive that I understand without problems.

[@rberdan1]

Hi Oliver very nice video - with phase as you said you really need a phase telescope to align the rings though you can also do this by looking down the eyepiece tube without the eyepiece in of course. Also instead of using a prepared slide, I think the best specimen you can use to demonstrate phase contrast is to use some cheek cells from the inside of your mouth and compare the image with bright field. Used Phase contrast is about $600 for the condenser and set of objectives from AM microscopes .For viewing pond microorganisms phase contrast really makes a big difference. I couldn't live without phase - I have all 4 versions of Olympus phase contrast objectives from negative to Positive Low Low, but one version is enough for most folks. I think the 10, 20 and 40X objectives are most useful, but the phase 100X oil does work for viewing smaller organelles like mitochondria.

[@johnf3668]

Robert, what "version" phase objectives would you recommend for "most folks". Thanks.

[@garyspencer1711]

Robert, I just started using my (used) phase contrast setup on (used) Olympus CX41 scope that I purchased last year. Purchased the CX-PCD from Toronto a few weeks ago for $249 US and I did not expect it to be in MINT condition. The darkfield illumination is much better than using the CH3-CD condenser with standard attachment lens (CX-AL) for 10x and 20x darkfield. If I use immersion oil on top of CH3-CD condenser, I can achieve fairly good 40x darkfield. Obviously, I have to move the condenser upward to come in contact with the bottom side of specimen slide. The Olympus CH3-CD is designed for immersion oils. I want to use immersion oil on my Phase Contrast turret condenser (only for 40x darkfield) but I can't find any specifications for the CX-PCD condenser. Even the Olympus user manual does not list ANY specs. Some people have claimed that any condenser that is not "marked" DRY, can be used with immersion oils. I don't trust these kinds of general statements .....yet. So, do you know if Olympus Phase Constrast turrets can be used with immersion oil ? Last topic: To align phase contrast condenser, I use a very old Canon camcorder looking down the eyepiece tube using camcorder manual focus adjust. This camcorder has a minimum focus distance of 2 cm ! Regards, Gary S.

[@johnharvey4496]

Get yourself an eyepiece "alignment telescope", a simple device to peer through the tube to check the alignment of the condenser annulus to the objective ring. You can cobble something up if you 'aint got one, but that's the way to setup for alignment so as to optimise your PC and minimise artefacts.

[@garbleduser]

I would like to see an extended version of this!

[@glasslinger]

I got my new moneypit (grin) an Olympus EH with the Olymups phase contrast set. I find the condenser different than yours. The adjustments on mine are on the stage rather than the condenser itself. And the index mark is on the right side instead of the front. I guess there were different models of phase contrast setups. Still having real trouble with the camera, trying to get on the screen as good as what is in the eyepiece. Looks like the moneypit is going to get deeper for a good camera.

[@johnharvey4496]

Love the enthusiasm, on the floor, down on your knees like a kid under the Christmas tree. Before that, open the box "newspaper!", I thought you were going to be distracted and start reading. It reminded me of the old days, where we would save string, newspaper and brown paper bags, back in the war days. Sorry I mentioned the war, gotta remember "Don't mention the war" :-)>

[@andrealucecita]

your channel is amazing im a chemistry student and i started some years ago to introduce myself in this world of microscopy, i love it i have learned a lot from you keep going

[@anbinhnguyen9395]

Very good explanation. You helped me understand the amplitude contrast vs Phase constrast.

[@Trylobyte]

I love phase contrast, finding it especially good for live protozoa and diatoms

[@DP-qb1zw]

I love it too. My default viewing of living samples

[@Microtonal_Cats]

I love this channel. I'm used to microscopy, this is helpful, and fun. More like a friend sitting next to you telling you how to do it. I dig my Michael Scope, and dig your work about michael scopes.

[@fonsrens5541]

At last !!! a very helpfull lesson this stirs me so i'm taking up again my books on optics. Thank you for your clear way of explanation ;

[@burtonkeeble670]

Hi Oliver, It is my understanding that phase contrast is often used in place of staining, in order to differentiate structures. My question: Can a phase contrast lens give you as good an image as a normal lens, in brightfield/darkfield? Or, would it just make sense to have a normal lens. My interest lies in subjects larger than bacteria, mostly protists. Would phase contrast give me enough improvement over normal images to justify the expense?

[@DP-qb1zw]

Yes, best for unstained but also works with faded, washed out stained slides and regular stained slides.

[@DP-qb1zw]

Theoretically, images have less resolution in bright field than regular objectives but I don't see a lot of difference when I compared them from the same company on the same microscope. I have one AO 10 microscope Ithat I can use for phase contrast, bright field, dark field and polarizing. And I sometimes combine PC or DF with polarizing.

[@johnf3668]

Awesome video Oliver! I learned a lot; I'm planning too add this capability to my American Optical 110. Thank you for all your great work!!

[@DP-qb1zw]

My AO 10 is setup with PC and gives outstanding views of living samples

[@VFXManiac]

I have a very old Meopta microscope. I had to make a few things by hand, such as lighting. Thanks to these imperfections, I can see objects in three dimensions, or as a phase contrast or darkfield. Just move the light source and use hand-made shades / filters/ circles with holes, etc .. Just experiments with light :)

[@lotharmayring6063]

DIY PH can easily made by soot from a candle and and scraping it off to give a stripe on both filters. A ring is not nessesary

[@bendirval3612]

Can you describe how you cleaned that objective? It seemed like the dirt was inside the objective itself.

[@geraldineward5657]

Being new at this study, I find your videos very helpful and interesting.

[@Mark-ks9jj]

Another great informative and easily understood video

[@emirselman3984]

yoooo, this was an incredible display and teaching. Thanks man! Definitely subscribing

[@jkallassy]

Great video - thanks! Do you have any thoughts on the quality of the relatively inexpensive phase contrast microscopes offered by Amscope (T490A-PCS or B390A-PCS)? I find it hard to believe that such feature-rich microscopes can cost less than $700 (camera additional).

[@cstephen98]

You refer to them as, 'rings' but would those be the prisms to first split and convert the light for viewing? Makes sense then you'd need one condensor per objective then so the prisms line up properly.

[@ThatFailedartist]

youre blowing my mind dude, please do a video on soil biology

[@jonathanvaucher2446]

Thanks for this! I just bought a Zeiss axioskop and it has a 0.9 Ph phase apochromatic aplanatic condenser with Ph and DIC turret (Zeiss 44 52 41) and one of my objective is a Plan-NEOFLUAR infinity 40x Ph2, but I'm clueless when it comes to phase contrast... So this vid is eye-opening

[@jonathanvaucher2446]

BTW, the prism in the trinocular head was misaligned because the glue (greyish substance) holding the prism to the metal bracket had cracked for drying... Any advice on what glue to use to secure it back to place ?

[@DP-qb1zw]

@@jonathanvaucher2446 Epoxy

[@lizweaver6467]

applaud this video however I was a bit appalled at the amount of dust etc. on the inside of the first objective. It wouldn't be seen as it would be out of the focal plane -most of it anyway. I just opened a well packed box of objectives I bought to find the sender just dropped all of them together in a bag without of even rolling each in paper. Will be inspecting them next. Hope microbehunter was able to clean his with just dry air.

[@shiwaz1594]

Congratulations on the upgrade, cant wait to see more samples! :)

[@claudelussier1678]

Look like this Phase Contrat set is a Nice Christmas gift to you!

[@keitarotamura493]

hello i have a question the block you add to the microscope i saw you have on amazon is it compatible to the swift sw380T if yes then i can order my order ? tks you

[@WillianMai]

I like your channel, but I think it would be better if you told when you are using 1000x and 400x and sometimes also telling some details on how you collected and which colorants you have used to show the images. It would help us to reproduce the same results for studying. My area is electronic engineering, nothing to do with microscopy, but I like and many people would love to know how to get the same results. It would make your channel more immersive for people that is learning. They would even get addicted i guess.

[@alanversfeld7497]

Thanks,I have bought an old Will Wetzlar ,just sorted out light (was missing ) I also await 100x object lens if you have advice happy to learn.

[@gauravmudgal841]

Solved my brainstorming with assembling the PCM I just got with me new. Thanks a lot. On my knees to u.

[@magnum127]

So if I use the phase contrast condensers but do not use the phase contrast objective, I then get the dark field picture right?😅😅

[@mohammadyaghoobi5740]

Great passion. I like the video and the explanation at the end

[@kislennox]

I'm just starting to understand phase contrast microscopy. As basically interested in photography, i found it very useful. Maybe a reall stupid question, but how do I know if the phase contrast set is compatible with my microscope? Thanks!

[@Microbehunter]

By talking to the manufacturer. The Phase Contrast objectives must be compatible to the condenser (phase ring and phase annulus must match) and then the condenser must be compatible to the microscope. Generally, if you want to buy phase contrast, it might be a better option to directly buy one complete, instead of upgrading an existing microscope. The costs are high anyway, so the savings that you get by upgrading might not be worth it (and there is a hassle putting a system together).

[@plabitapaul5696]

Thanks a lot sir... Easy to remember and understand... 🤩

[@CAKRAWALAMIKROSKOP2021]

agree

[@michellec4511]

Thank you!! I really enjoyed your explanations!

[@VersaiOnline]

... so why did you show us dead stained slides if phase contrast is for live specimens?

[@flugschulerfluglehrer]

Some spores have a relief on their surface. These spores are typically less than 10um. So these structures are close to the resolution of a good 100x objective. Would phase contrast make them sharper or blurrier? The exact details are sometimes crucial to correctly determine the species you have on the slide.

[@DP-qb1zw]

Oblique illumination might greatly increase your resolution

[@flugschulerfluglehrer]

@@DP-qb1zw Thank you for the tip. Looks promising. I’ll definitely give them a try.

[@kjellgolsen8521]

Can one use phase contrast objectives and do darkfield ?

[@Microbehunter]

Yes.

[@das250250]

Isn't the brightness proportional to the number of photons making it through( being reemitted from ) the specimen in bright contrast rather than intensity. Intensity to the eye is about number of photons not amplitude

[@johnanderson1254]

I have a question Oliver. I have a phase contrast inverted microscope I just recently purchased from my university. It required quite a bit of work to get it back in working condition but it includes a GIF (green interference filter) what is the purpose of this filter and is it necessary to use phase contrast? When I use the filter it merely turns everything green.

[@Microbehunter]

I do not have this green filter and it also works. Theoretically the contrast should be a bit lower without the filter. The optics have been designed to work best in monochromatic green light. You can not take all wavelengths into consideration, so the optics were calibrated to work best for green light, an intermediate wavelength. So if you want to get the theoretically best contrast, you should use the green filter. But is it not necessary and apparently with modern objectives the difference in contrast can be ignored.

[@mazyar_]

Hello Oliver, Under brightfield, are there image contrast differences between a "PL" and a "non-PL" objective of the same magnification? Thank you.

[@Microbehunter]

Yes, there is a small difference, but the difference is small. I made a video about this topic some time ago: kzbin.info/www/bejne/iXy9f4Fqg5mlf5I

[@mazyar_]

@@Microbehunter Thank you.

[@santanadas8137]

Sir, how to take picture from this? Please help me out

[@garbleduser]

I have found that some endoscope cameras can lock onto microscope eyepieces.

[@wolfram77]

I cant wait to buy a micoroscope too, insprired by you. i want to see yeast under a microscope. just today the bottle i was keeping for growing natural yeast became active and i wish to use it to make sourdough bread. thanks.

[@chairulanwar6093]

hai, recently i got secondhand olympus imt2 phase contrast for IDR 500.000,or for abaut 50 USD, the lamp is working, the lens its complete, but i cannot see any object in my sample glass, where i can get any advice for my problem?, thank you so much

[@ceilingfun2182]

Can you upgrade swift SW380t to phase contrast?

[@Itshamsul]

Sample preparation in phase contrast microscopy? Any video?

[@astro-maths2362]

17:20 3, 2, 1 Phase Contrast !!! 😃👍

[@srigirivasramakrishnan96]

You are really good. Really really good.

[@himasuchiang8842]

Wow thanks really need this

[@youknowitis]

Very helpful thanks man Nice explanation 👍👍

[@donaldeisenbarth5255]

I, being use to bright field, was not impressed with phase contrast. Not worth the expense for most. And I agree the tri ocular is great as I use a simple Nikon camera and the self focusing feature results in excellent pictures. The camera is light enough I can sit it on top of the objective without support.

[@DP-qb1zw]

I started with bright field and then got PC setup which I enjoy very much on living samples.

[@SibiriDavid]

that is very understandable. well done🤭👏

[@powertube5671]

Nice Job! Thank you!

[@Welch_Marketing]

I have a Nikon e400. Supposedly the objectives standards allow for phase contrast. Do i just need a phase contrast condenser?

[@Microbehunter]

You need phase contrast objectives (they are not normal objectives) and the correct phase contrast condensor. They have to match.

[@PredatorGaming-2007]

Can you explain what is a phase contrast??? I didn't understand well

[@johnharvey4496]

Last part of the video - explained reasonably well

[@manudehanoi]

u didnt show setting up the height of the condenser

[@johnharvey4496]

True, need to get that right first up. Do the whole setup many times and it becomes second nature. It's worth the trouble but.

[@totoff92]

hello, do you have to be absolutely in Kohler illumination (as you didnt mention that you set it before) ? My microscope cant be in kohler so i wanted to know. Thanks

[@DP-qb1zw]

No

[@finpark7785]

I was hoping you could tell my what sort of Phase Contrast Condenser I would need for a Motic B310. Or are they all compatible with one another. If I were to pick up a Nikon Phase Contrast Condensers, would it work on a B310?

[@Microbehunter]

The Phase Contrast Condensers are not generally compatible. They must fit into the microscope and also must be optically compatible with the objectives. Contact Motic directly and ask for an offer. From the Motic website we know that the B310LED series allows for Phase Contrast, so it should be possible in principle to upgrade the microscope.

[@finpark7785]

@@Microbehunter What is the easiest way to experience Darkfield/bright field? is there a way to just add a filter? I love your channel. I'm trying to learn as much as I can from you.

[@DP-qb1zw]

@@finpark7785 I've experimented with making patch stops for DF by making different size black ⚫ circles on a glass slide and putting it on my filter holder to block the light. You can also get oblique illumination that way.

[@lotharmayring6063]

Es ist nicht wahr, dass man Unterschiede im Brechungsindex in der Hellfeldmikroskopie nicht sieht oder darstellen kann. Dazu muss man lediglich den Einfallswinkel des Beleugtungsstrahls entsprechend veraendern. Ich sehe wenig Unterschiede von Phasenkontrast zu einer Kombination aus Hellfeld, Dunkelfeld und Schraegbeleuchtung die den extremen nicht gerechtfertigten finanziellen, apperativen und Einstellaufwand zumindest fuer den Amateurmikroskopiker rechtfertigen wuerden, auch wenn man gebraucht kauft. Unterschiede im Brechungindex von Objekten lassen sich durch variable schraege Beleuchtung im Hobbybereich genuegend befriedigend darstellen. Ich habe auch nie verstanden warum es nicht moeglich ist den Phasenring ausserhalb von Objetiven zu plazieren. Auch der Phasenkontrastkondensor besteht ja nur aus Ringblenden die man selbst anfertigen und einlegen kann. Die 1000 Euro sollte man lieber in einer guten Beleuchtung, Kondensor, Objektiven und Okularen anlegen. So for Hobbyists phasecontrast is bullshit.

[@Al1ceBajenova]

TY! Veru usefull video!

[@edward_grabczewski]

Some objectives have letters before the magnification number. For example, here in this video I can see "PL10" and on one of my objectives it reads "S40". What does the "PL" and "S" mean?

[@Microbehunter]

"S" I don't know. PL stands for "positive low" it is probably a phase contrast objective. You can easily find this out by taking it out and looking though it from the back. If you can see a ring on the lens, then it is a phase contrast objective. Might be a manufacturer specific designation. So if you know the make of the objective, one could check with the manufacturer.

[@edward_grabczewski]

@@Microbehunter I've found out that "S" means "Spring mounted" and it means the objective lens closest to the slide is spring loaded, to avoid damaging the lens or slide.

[@Microbehunter]

Very good. Thank you for the information. Interesting.

[@edward_grabczewski]

@@Microbehunter I checked my S40 lens and yes, it moves up and down on a spring if you press it. The others don't. I'm trying to find an S60 lens in the same series but all I can see are straight 60X lenses by the same manufacturer. Presumably they don't move, but I've sent out a query to the seller to check. The objectives on my Swift SW150 and AmScope M150 are identical and cheap, but quite acceptable quality. These microscopes are for my children. I might buy a better one for myself. The AmScope T120C or T490 seem to have all the features I need for now. The manual for the T490 series is worth reading!

[@geometricart7851]

I would just make your own darkfield filter with no need for phase contrast

[@DP-qb1zw]

Dark field and phase contrast are 2 vastly different things.

[@vallos3]

Good Video, but i have to make one point. Light does not have an amplitude, the brightness comes from the number of photons. As soon as a Photon is detected it is destroyed completely. It cannot lose parts of its energy, its energy is determined by its wavelength. The basic principle still consists. G'day

[@psynurse]

you are incorrect. Light also acts as a wave therefore does amplitude

[@johnharvey4496]

Let's discuss the particle-wave theory

[@vallos3]

@@johnharvey4496 Go on then :) The frequency of the light is its energy and thus it's colour.

[@kambe9713]

Very helpful. Thank you

[@rishabhanand9751]

Where is the role of phase ring? You only showed 90 degree phase shift that will not create interference.

[@andregomesdasilva]

Great video

[@Alpha-Quantum-kg9pe]

Thank you,

[@Martinko_Pcik]

Does the condenser and objective need to be from the same manufacturer for phase contrast ?

[@Microbehunter]

Yes, the phase ring in the objective and the phase annulus in the condenser must match in size. Even different generations of objectives/condensers from the same manufacturer might not fit together.

[@johnharvey4496]

Kinda, yes. But its not so bad as iphone vs android. You're rolling the dice if you select them separately or from different manufacturers.

[@DP-qb1zw]

Most of the time. I have success using a Nikon condenser with Zeiss phase objectives on a Nikon L-KE microscope. The the 10x uses the 10 ring and the 16x and 40x both use the 40 ring.

[@rachelgrace6708]

16:20 theoretical explanation.

[@azharmohd6930]

Link please

[@PredatorGaming-2007]

Thanks. Can we make own phase contrast????????

[@Microbehunter]

No. You need a separate phase contrast objective with a phase ring, and it is not possible to put the phase ring on the lens of the objective yourself. You could make the phase annuli yourself for the condenser, though.

[@PredatorGaming-2007]

@@Microbehunter thanks but I made my own and I keep it near the condenser. It works good

[@Microbehunter]

@@PredatorGaming-2007 Cool! How did you put the phase ring on the objective lens? Or did you use some foil?

[@PredatorGaming-2007]

@@Microbehunter No. I use a compound microscope. In its condenser, there is a ring. I think we make our own phase contrast using plastic sheet. we can put it there in the platform.

[@johnharvey4496]

To a certain extent, yes. Make your own condenser as you say, but best to buy the objectives with the ring built it because it's so small to get into a regular objective.

[@o.k.6322]

Hi. Thanks. Very interested. Can you see coronavirus in optical microscope . Can't find...

[@DP-qb1zw]

No

[@o.k.6322]

@@DP-qb1zw 🤣🤣🤣🤣 thanks . I'm sorry .but after 2 years (sins asck)that not actual...

[@boris2342]

fingerprints in phase cont ???

[@samansabetghadamhaghighi9143]

Awesome

[@azharmohd6930]

Pirchase link please

[@Microbehunter]

This was bought second hand. You have to check eBay. The condenser will only work with specific Olympus models and objectives.

[@MadonnaPaul-qd8sf]

Soul exchange

[@vaccinefraud5570]

I've been trying to catch up with your EXCELLENT presentations (you are a Geek's GEEK) while trying to select a scope that has multiple features from brightfield to phase contrast to darkfield BUT EVEN THE EXPERTS couldn't explain to me how to get all three in one unit. Using the different anuli on the to create the occultation for the darkfield at different objectives makes PERFECT SENSE, YET NO ONE IN THE INDUSTRY COULD EXPLAIN THAT TO ME! My question then would be if all phase contrast condensers are similar to the one you are featuring so that we can all have MULTIFUNCTION devices just like you? I wish I could apply INFINITE THUMBS UP TO YOUR VIDEOS!

[@johnharvey4496]

If you buy a decent PC condenser, it will come with darkfield and brightfield positions