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 Post subject: Darkfield photography
PostPosted: Sat Oct 06, 2018 2:18 am 
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My new oiled darkfield condenser is giving great views in the eyepiece but not on the Canon 60Da DSLR camera in the trinoc. port. The camera images seem blurred and the colours are strange. I’ve tried a variety of settings of Tv,P,Av and speeds but all the results have been disappointing. I’m including a cropped image as an example.

Are there any recommendations about how to get better results? Thanks

Michael


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PostPosted: Sat Oct 06, 2018 3:18 am 
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I'm wondering if a combination of a somewhat long exposure and camera shake is the problem? If so, might want to mount the camera independently of the microscope. Are there settings for your camera to reduce shutter bounce, such as a live view setting?


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PostPosted: Sat Oct 06, 2018 3:36 am 
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How are you attaching the camera to the trinoc port?

How are you focusing?

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PostPosted: Sat Oct 06, 2018 3:51 am 
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What ISO setting did you use? The image looks as if it were taken with a very high ISO setting. Don't use Auto-ISO but set the camera fixed on the lowest ISO setting like 100.

Many Canon cameras can be released from live view without the shutter or mirror moving before the exposure. Does your camera offers this electronic first shutter curtain feature and did you use it?


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PostPosted: Sat Oct 06, 2018 7:25 am 
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What is the actual size of the object and with which optics (mag, dry/oil, etc) was is imaged? by how much is it cropped?

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PostPosted: Tue Oct 16, 2018 9:18 pm 
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MicroBob wrote:
What ISO setting did you use?
Don't use Auto-ISO but set the camera fixed on the lowest ISO setting like 100.


I agree, but not (low) if the camera doesn't have 100, as low is no lower than 200 but uses a sort of emulation which can cause issues.


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PostPosted: Wed Oct 17, 2018 2:13 pm 
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DF is partially a product of refraction, which causes chroma. Peering into transparent structures, illuminated by an oblique light source, with high magnification will show some refracted chromatic aberration. Very often, your eye doesn't see it as much or not at all but the camera will record it. It becomes more apparent in a crop.

There are a number of oil immersion DF condenser designs. As the design gets more precise and expensive, the freedom from ca does too. The best are ca free mirror condensers, such as cardioids; a commonly available choice, with paraboloids a little lower on the totem pole and stops in the various lens condensers further down depending on the achromatism of the condenser.

The difference between achromats and semi and full apochromat objectives also becomes more apparent with DF.

A further complication is just how well the photo relay lens is matched to the objective in question. In the past, I have had a situation where I was using a relay lens on a certain microscope that based on it's results using achromats, seemed very good. It turned out to be almost a perfect match but not quite. When used with apos and especially with DF, chroma would show up where I did not expect it. After a period of head scratching , I decided that it was the relay lens and went on a search for a replacement, which solved a lot of the chroma problem. There did not seem to be much improvement in BF or phase, especially with achromats as in DF with apos. DF is essentially a black and white image, so colour variations are more defined and more apparent.


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PostPosted: Wed Oct 17, 2018 6:24 pm 
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apochronaut wrote:
DF is partially a product of refraction, which causes chroma...DF is essentially a black and white image, so colour variations are more defined and more apparent.
One way to get rid of the CA and perhaps improve the image would be to use a narrow bandpass filter, that transmits light within a narrow wavelength range only. Interference filters are best, but they are expensive. I would try with a 40-60nm filter, say, a blue - 430-470nm or green (530-570nm) filter, or give it a try with an ordinary green or blue filter, it might improve the image by removing most of the CA. Alternatively, I would consider a LED lamp, not white but more monochromatic.

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PostPosted: Wed Oct 17, 2018 6:54 pm 
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I think we really need to know how you are attaching the camera first. I have used a Canon 60D dSLR and had no issues with dark field like this. I used an adapter that attaches to your camera like a normal lens and fits into eye pieces or C-mounts. $60 on amazon. Something like this, but I bought it when it was cheaper:

https://www.amazon.com/OMAX-Microscope- ... pe+adapter

mnmyco


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PostPosted: Wed Oct 17, 2018 7:00 pm 
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I haven't had much luck with led for DF, even when one can get enough light out of them. I am using apos, and the narrow wavelength they provide, limits the information available. I'm not a fan of the heavy blue cast to the image, either. Perhaps that would be of value in reducing extraneous information, with a chromatic aberration prone system.
DF is hard enough to squeeze natural colour from anyway, and leds just make matters worse. Ditto for filtering.

To boot, there is enough negative literature out there about them, that I am cautious about their safety in a blast furnace lighting sort of situation , like a microscope field. They don't even make decent Christmas lights.


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PostPosted: Sun Oct 21, 2018 3:54 pm 
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apochronaut wrote:
I haven't had much luck with led for DF, even when one can get enough light out of them. I am using apos, and the narrow wavelength they provide, limits the information available. I'm not a fan of the heavy blue cast to the image, either. Perhaps that would be of value in reducing extraneous information, with a chromatic aberration prone system.
DF is hard enough to squeeze natural colour from anyway, and leds just make matters worse. Ditto for filtering.

To boot, there is enough negative literature out there about them, that I am cautious about their safety in a blast furnace lighting sort of situation , like a microscope field. They don't even make decent Christmas lights.


We have been through all of this LED crap several times over the years... You know yourself that the big makers use LED lighting in their top of the line stands, yet you continue to talk against them.. Do you just want to stay in the olden times, or can you open your mind and follow new processes???....

https://www.leica-microsystems.com/prod ... dm-il-led/


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PostPosted: Sun Oct 21, 2018 4:27 pm 
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billbillt wrote:
apochronaut wrote:
I haven't had much luck with led for DF, even when one can get enough light out of them. I am using apos, and the narrow wavelength they provide, limits the information available. I'm not a fan of the heavy blue cast to the image, either. Perhaps that would be of value in reducing extraneous information, with a chromatic aberration prone system.
DF is hard enough to squeeze natural colour from anyway, and leds just make matters worse. Ditto for filtering.

To boot, there is enough negative literature out there about them, that I am cautious about their safety in a blast furnace lighting sort of situation , like a microscope field. They don't even make decent Christmas lights.


We have been through all of this LED crap several times over the years... You know yourself that the big makers use LED lighting in their top of the line stands, yet you continue to talk against them.. Do you just want to stay in the olden times, or can you open your mind and follow new processes???....

https://www.leica-microsystems.com/prod ... dm-il-led/
I think one should distinguish between the big makers and hobby microscopysts here, where "hobby" refer to folks that convert their illumination to LED under limited budget contraints, and aim mainly at color photography.
billbillt, the link you posted sets a perfect example. Leica promotes a cell culture microscope, and I believe that a large portion of their market is cell biology microscopy. the leading modality in biology today is fluorescence, due to various reasons. Confocal microscopy is based on lasers anyway. But fluorescence can be done with LEDs as well, since they are fairly monochromatic. So they are fine for excitation. Likewise, DIC and phase can be done successfully with LEDs, when the result is gray scale or pseudo-color. Many research-data images today are "colored" by software anyway. Thus, for up-to-date research, LEDs are fine, and not white light LEDs but especially blue, green, LEDs etc. And these LEDs are quite expensive - hundreds of USD each, for the single wavelength/color.
On the other hand, for us folks who would like just to upgrade an old hobby microscope from the obsolete incandescent or the rare to find halogen lamp, and produce pretty color images, I agree with most of what Apochronaut says, although I am less concerned about UV emission.

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PostPosted: Sun Oct 21, 2018 4:41 pm 
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Hobbyst46 wrote:
billbillt wrote:
apochronaut wrote:
I haven't had much luck with led for DF, even when one can get enough light out of them. I am using apos, and the narrow wavelength they provide, limits the information available. I'm not a fan of the heavy blue cast to the image, either. Perhaps that would be of value in reducing extraneous information, with a chromatic aberration prone system.
DF is hard enough to squeeze natural colour from anyway, and leds just make matters worse. Ditto for filtering.

To boot, there is enough negative literature out there about them, that I am cautious about their safety in a blast furnace lighting sort of situation , like a microscope field. They don't even make decent Christmas lights.


We have been through all of this LED crap several times over the years... You know yourself that the big makers use LED lighting in their top of the line stands, yet you continue to talk against them.. Do you just want to stay in the olden times, or can you open your mind and follow new processes???....

https://www.leica-microsystems.com/prod ... dm-il-led/
I think one should distinguish between the big makers and hobby microscopysts here, where "hobby" refer to folks that convert their illumination to LED under limited budget contraints, and aim mainly at color photography.
billbillt, the link you posted sets a perfect example. Leica promotes a cell culture microscope, and I believe that a large portion of their market is cell biology microscopy. the leading modality in biology today is fluorescence, due to various reasons. Confocal microscopy is based on lasers anyway. But fluorescence can be done with LEDs as well, since they are fairly monochromatic. So they are fine for excitation. Likewise, DIC and phase can be done successfully with LEDs, when the result is gray scale or pseudo-color. Many research-data images today are "colored" by software anyway. Thus, for up-to-date research, LEDs are fine, and not white light LEDs but especially blue, green, LEDs etc. And these LEDs are quite expensive - hundreds of USD each, for the single wavelength/color.
On the other hand, for us folks who would like just to upgrade an old hobby microscope from the obsolete incandescent or the rare to find halogen lamp, and produce pretty color images, I agree with most of what Apochronaut says, although I am less concerned about UV emission.


DID YOU BOTHER TO SCROLL THROUGH ALL OF THE OFFERINGS FROM THEM??... IT IS NOT JUST "CELL CULTURE MICROSCOPES" THAT LEDS ARE USED IN... ZEISS DOES IT ALSO.. DO I HAVE TO FIND THAT AND POST A LINK ALSO??... DON'T YOU THINK THESE REPUTABLE MFGS HAVE NOT SOLVED THE COLOR PROBLEMS HERE??.. LEDS ARE THE FUTURE OF MICROSCOPE LIGHTING... I ALSO SAW A WHILE BACK LEDS ARE BEING USED IN FLUORESCENCE MICROSCOPY... MOST OF THIS OBJECTION IS JUST A RESISTANCE TO CHANGE...


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PostPosted: Sun Oct 21, 2018 6:13 pm 
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billbillt wrote:
DID YOU BOTHER TO SCROLL THROUGH ALL OF THE OFFERINGS FROM THEM??... IT IS NOT JUST "CELL CULTURE MICROSCOPES" THAT LEDS ARE USED IN... ZEISS DOES IT ALSO.. DO I HAVE TO FIND THAT AND POST A LINK ALSO??... DON'T YOU THINK THESE REPUTABLE MFGS HAVE NOT SOLVED THE COLOR PROBLEMS HERE??.. LEDS ARE THE FUTURE OF MICROSCOPE LIGHTING... I ALSO SAW A WHILE BACK LEDS ARE BEING USED IN FLUORESCENCE MICROSCOPY... MOST OF THIS OBJECTION IS JUST A RESISTANCE TO CHANGE...
I do not see objections, only controlled reservations.

I just opened the (current) Zeiss site:

https://www.zeiss.com/microscopy/us/sol ... scopy.html

And citing from it:
"Among the most promising of emerging technologies for illumination in optical microscopy is the light-emitting diode (LED). These versatile semiconductor devices possess all of the desirable features that incandescent (tungsten halogen) and arc lamps lack, and are now efficient enough to be powered by low-voltage batteries or relatively inexpensive switchable power supplies. The diverse spectral output afforded by LEDs makes it possible to select an individual diode light source to supply the optimum excitation wavelength band for fluorophores spanning the ultraviolet, visible, and near-infrared regions. Furthermore, newer high-power LEDs generate sufficient intensity to provide a useful illumination source for a wide spectrum of applications in fluorescence microscopy, including the examination of fixed cells and tissues, as well as live-cell imaging coupled to Förster resonance energy transfer (FRET) and lifetime measurement (FLIM) techniques. The interactive tutorial featured in this section explores the ZEISS Colibri LED illumination system for widefield fluorescence microscopy." ALL LED applications that they talk about are FLUORESCENCE.

Then I opened another Zeiss site, about students microscopes: they sell the Primo Star, and here is a citation:

"Clever Details for More Freedom
Choose between 30 W halogen or energy saving LED illumination.
In areas with fluctuating or no electricity you profit from the battery supply unit.
Upgrade Primo Star with the fluorescence tube to get a LED fluorescence microscope."


Please note, that there is no real dispute between us, and I hope nobody is upset. The big makers provide LED sources, and I am convinced that you can easily prove it with examples. These LEDs are not cheap - have purchased and used them. The only point is that are not necessarily the best solution for hobby use. I am the fortunate/unfortunate user of a relatively inexpensive LED light...

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PostPosted: Sun Oct 21, 2018 6:28 pm 
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I DON'T THINK THAT THEY ARE EXPENSIVE... THAT IS A MYTH.... JUST PAYING MORE FOR SOMETHING DOES NOT MEAN IT IS BETTER... YOU CAN WORK WITHIN THIS CHART AND DO JUST ABOUT ALL YOU NEED TO...


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PostPosted: Sun Oct 21, 2018 9:57 pm 
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billbillt wrote:
... YOU CAN WORK WITHIN THIS CHART AND DO JUST ABOUT ALL YOU NEED TO...

Sorry, billbillt ... I have to disagree with that assertion
The lower part of the chart is fine, but the first two rows conceal a problem.

Please have a look at this previous thread: http://www.microbehunter.com/microscopy-forum/viewtopic.php?f=9&t=5686 where Hobbyst46 and I discussed 'White' LEDs.

I suspect that, in common with many others, apochronaut sees tungsten filament lighting as the definitive baseline for microscope illumination ... and although they are very useful 'White' LEDs remain a poor approximation to that [Black Body Radiation].

Of the two LED types; Warm White has the better CRI [colour rendering index] but its spectral distribution is still very peaky.

MichaelG.

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PostPosted: Sun Oct 21, 2018 10:54 pm 
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I USE WARM WHITE BECAUSE IT LOOKS LIKE NATURAL SUNLIGHT..... I POSTED THIS CHART SO A PERSON COULD SELECT AN LED WITHIN THE CHOSEN WAVELENGTH.... HIGH QUALITY STANDS THAT COME WITH LED LIGHTING WORK FINE..... IT IS JUST NOT RIGHT TO DEMAND THAT A PERSON USE SUCH AND SUCH LIGHTING, WITH SUCH AND SUCH STANDS OR IT CAN'T BE ANY GOOD... I LIKE TO EXPERIMENT WITH THINGS.. THAT WAS MY JOB AS AN ENGINEER...KOHLER ILLUMINATION IS NOT A REQUIREMENT FOR GOOD MICROSCOPY... IT IS NOT EVEN NECESSARY WITH LED LIGHTING, AS YOU HAVE NO FILAMENT TO WORRY ABOUT... I SEE A LOT OF THIS AS NOT BEING COMPLETELY NECESSARY FOR HOBBY WORK.....


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PostPosted: Sun Oct 21, 2018 11:32 pm 
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@billbillt Kohler also removes the appearance of dust on parts of the optical system and removes unnecessary light. So, it does more than just remove the filament image.

If I recall correctly, the only issue with LED I am aware of is that you cannot use a simple rheostat to control brightness without having temp changes in the light.

I have personally used a Canon 60D through a trinocular head on an orthulux with an LED on a heatsink taped to the lamp housing with DF and had no issues with photos.

mnmyco


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PostPosted: Sun Oct 21, 2018 11:45 pm 
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mnmyco wrote:
@billbillt Kohler also removes the appearance of dust on parts of the optical system and removes unnecessary light. So, it does more than just remove the filament image.

If I recall correctly, the only issue with LED I am aware of is that you cannot use a simple rheostat to control brightness without having temp changes in the light.

I have personally used a Canon 60D through a trinocular head on an orthulux with an LED on a heatsink taped to the lamp housing with DF and had no issues with photos.

mnmyco


NOW YOU ARE TALKING.... I HAVE HAD GOOD LUCK WITH THEM ALSO.... LEDS ARE CONTROLLED BY CHANGING THE FORWARD CURRENT, NOT VOLTAGE... GLAD YOU HAD GOOD LUCK...


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PostPosted: Mon Oct 22, 2018 4:53 am 
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billbillt wrote:
... I SEE A LOT OF THIS AS NOT BEING COMPLETELY NECESSARY FOR HOBBY WORK.....

Now that ^^^ is a much better way of putting it.

MichaelG.

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PostPosted: Mon Oct 22, 2018 1:34 pm 
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billbillt wrote:
apochronaut wrote:
I haven't had much luck with led for DF, even when one can get enough light out of them. I am using apos, and the narrow wavelength they provide, limits the information available. I'm not a fan of the heavy blue cast to the image, either. Perhaps that would be of value in reducing extraneous information, with a chromatic aberration prone system.
DF is hard enough to squeeze natural colour from anyway, and leds just make matters worse. Ditto for filtering.

To boot, there is enough negative literature out there about them, that I am cautious about their safety in a blast furnace lighting sort of situation , like a microscope field. They don't even make decent Christmas lights.


We have been through all of this LED crap several times over the years... You know yourself that the big makers use LED lighting in their top of the line stands, yet you continue to talk against them.. Do you just want to stay in the olden times, or can you open your mind and follow new processes???....

https://www.leica-microsystems.com/prod ... dm-il-led/


The led systems used by Leica are highly engineered systems and are offered as an option. It is unlikely that cheapo led systems are going to provide the quality of imaging many skilled microscope users expect. Most of the sales of better professional grade microscopes include halogen lighting as a matter of course. It's not that I haven't tried led illumination systems, it's that they fall short in performance and they also may be dangerous, especially to children. Since you are so adamant that led is a fine illumination source, how about sending us some pictures of the quality of work I should be expecting.


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PostPosted: Mon Oct 22, 2018 1:53 pm 
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The led systems used by Leica are highly engineered systems and are offered as an option. It is unlikely that cheapo led systems are going to provide the quality of imaging many skilled microscope users expect. Most of the sales of better professional grade microscopes include halogen lighting as a matter of course. It's not that I haven't tried led illumination systems, it's that they fall short in performance and they also may be dangerous, especially to children. Since you are so adamant that led is a fine illumination source, how about sending us some pictures of the quality of work I should be expecting.

YOU DON'T KNOW YOUR BUTT FROM A HOLE IN THE GROUND.. STOP BEING A GROUCHY OLD MAN AND LET FOLKS EXPERIMENT WITH WHAT THEY WANT...


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PostPosted: Mon Oct 22, 2018 2:29 pm 
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billbillt wrote:
The led systems used by Leica are highly engineered systems and are offered as an option. It is unlikely that cheapo led systems are going to provide the quality of imaging many skilled microscope users expect. Most of the sales of better professional grade microscopes include halogen lighting as a matter of course. It's not that I haven't tried led illumination systems, it's that they fall short in performance and they also may be dangerous, especially to children. Since you are so adamant that led is a fine illumination source, how about sending us some pictures of the quality of work I should be expecting.

YOU DON'T KNOW YOUR BUTT FROM A HOLE IN THE GROUND.. STOP BEING A GROUCHY OLD MAN AND LET FOLKS EXPERIMENT WITH WHAT THEY WANT...


YOU SENT ZERO PICTURES... YOU CAN'T EVEN DO THAT WITHOUT A FAIL.... YOU WHINE ABOUT PHOTO EDITING OF PICTURES HERE ONLY BECAUSE IT IS BEYOND YOUR CAPABILITY TO DO IT... ON SECOND THOUGHT HERE, I AM GOING TO TO SET YOU TO "FOE", SO I NO LONGER HAVE TO ENDURE YOUR PREPOSTEROUS CRAP...


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PostPosted: Mon Oct 22, 2018 2:36 pm 
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Quote:
I think we really need to know how you are attaching the camera first.
If you can post some photos of your setup I think it will help.
Quote:
Don't use Auto-ISO but set the camera fixed on the lowest ISO setting like 100.
Agree on the need to keep ISO as low as possible.

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PostPosted: Mon Oct 22, 2018 2:39 pm 
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Quote:
YOU DON'T KNOW YOUR BUTT FROM A HOLE IN THE GROUND.. STOP BEING A GROUCHY OLD MAN AND LET FOLKS EXPERIMENT WITH WHAT THEY WANT...



Whoa, let's just simmer down. There is no reason for ad hominem attacks. There is plenty of room regarding this subject (LED lighting) for personal taste and opinions, without getting one's knickers in a twist.

There is no doubt that LEDs are the wave of the near-future, but right now they are not the solution for everything in all cases, otherwise there wouldn't be a filament bulb left on the planet. At the hobby level, a lot has to do with personal preference, economics, technical feasibility, and other reasons. It is up to each individual to make a decision for themselves as to what they want to do, and if the results satisfy them. Since it's a hobby, everyone has the complete right to do as they wish. But nonetheless, other viewpoints are valuable because they feed into one's knowledge base for making decisions.

I hope we can get back to MikeBradley's question.


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PostPosted: Mon Oct 22, 2018 2:58 pm 
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wporter wrote:
Quote:
YOU DON'T KNOW YOUR BUTT FROM A HOLE IN THE GROUND.. STOP BEING A GROUCHY OLD MAN AND LET FOLKS EXPERIMENT WITH WHAT THEY WANT...



Whoa, let's just simmer down. There is no reason for ad hominem attacks. There is plenty of room regarding this subject (LED lighting) for personal taste and opinions, without getting one's knickers in a twist.

There is no doubt that LEDs are the wave of the near-future, but right now they are not the solution for everything in all cases, otherwise there wouldn't be a filament bulb left on the planet. At the hobby level, a lot has to do with personal preference, economics, technical feasibility, and other reasons. It is up to each individual to make a decision for themselves as to what they want to do, and if the results satisfy them. Since it's a hobby, everyone has the complete right to do as they wish. But nonetheless, other viewpoints are valuable because they feed into one's knowledge base for making decisions.

I hope we can get back to MikeBradley's question.



I agree 100%.. And we can also stop trying to dominate and command what other folks want to do...

BillT


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PostPosted: Mon Oct 22, 2018 3:14 pm 
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All I asked ; was for you to send some pictures of what I could expect. I have no idea why , you are bursting a seam, Billbilt.


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PostPosted: Wed Mar 06, 2019 11:45 pm 
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Interesting picture
But it has a bad quality
Maybe it's your skills?
This site will help you improve them
http://fixthephoto.com/blog/photo-tips/ ... nners.html

Good luck


Last edited by itsnotyou on Thu Mar 14, 2019 3:52 pm, edited 1 time in total.

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PostPosted: Thu Mar 07, 2019 12:55 am 
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It's worth mentioning that the 60D disappointed a lot of high mag micro folk because it DOES vibrate when the electronic first shutter curtain "opens". My 80D does it too, though I think somewhat less.

If a continuous light exposure is long enough (say 2secs + ) then the vibration is only a small part of the exposure so it doesn't seem to be noticeable.


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