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Confocal microscope aligment

I’m using classic confocal microscope system. The laser wavelength is $405$ $nm$ and the objective is $80x$, $Na=0.75$, $\infty /0$. The pinhole diameter is $15 $ $\mu m$. What should be the precision ...
Malum Phobos's user avatar
4 votes
3 answers
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Is a subnucleus microscope really possible?

The reason mankind does not now have a sub-picometre microscope is generally attributed to the inability to create a perfect optical system. The question is, is this true? Consider a perfect optical ...
Ember Edison's user avatar
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Nyquist criteria along longitudinal axis

I understand that magnification of an imaging system must be so chosen so that at least 2 pixels sample the best lateral resolution (x-y) as per the Nyquist criteria. So in optical microscopy, if we ...
user1229009's user avatar
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1 answer
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de Broglie wavelength and resolution of microscope

I read about electron microscopes and it's said that electrons have a smaller deBroglie wavelength than the wavelength of light and therefore, better spatial resolution (lambda/numerical aperture). ...
user1229009's user avatar
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How electrons are produced in Scanning Electron Microscopy (SEM)?

I am referring to thermionic emitter using tungsten filament, as a current is applied to the filament and transfer heat until sufficient energy overcomes the work function, then the electron are ...
Shira's user avatar
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What happens if a pulsed laser beam is directly incident on the objective lens of a microscope?

I am currently building a microscope in my lab. In my setup, the pulsed laser beam is directly incident on the objective lens, rather than the typical configuration where collimated light is incident ...
RUYI ZHAO's user avatar
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Area Cross-Section of a dye / Trying to understand the interaction between photons and fluorophores

What is the right way to research / measure the cross-section of fluorophores and compare this to excitation illumination to arrive at the probability of an interaction and hence an emission? ...
user3533030's user avatar
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1 answer
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Shot noise limited detection

can somebody please explain the significance of shot noise limit? I am trying to understand why and how shot noise degrades image quality, for e.g., while imaging weakly scattering specimens using an ...
user1229009's user avatar
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Is it possible to determine the magnification of a compound microscope given the objective focal length, eyepiece focal length, and objective di?

Question as stated above, I want to know if it is possible to determine the magnification of the microscope from the objective and eyepiece focal lengths, and image distance of the objective lens. ...
Astrovis's user avatar
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Does the coherence of light sources make a difference in exciting fluorescence?

Using LED vs laser as light sources in wide-field epi-fluorescence microscopy are both popular choices. It seems that high intensity single color LEDs (such as this one) emits high radiometric power ...
Jason M's user avatar
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Electron microscopy and Interatomic distances of miller planes

I am studying Transmission Electron Microscopy (TEM), and have been seeing in articles TEM images of different materials typically come accompanied by these diffraction patterns, caused by the ...
Rye's user avatar
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Can we use prism in fluorescence microscope instead of dichroic mirror?

For the light path outlined here, if we use a prism instead of a dichroic mirror, the excitation light can also pass through the prism, but the emission filter can block the excitation wavelength if ...
nancy M's user avatar
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2 answers
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Why is the shorter the wavelength, the smaller the object you can image? and vice-versa?

I know that the shorter the wavelength the smaller the object you can image clearly. Why wavelength matters in imaging something? How having big wavelength wont let u image smaller object, like if u ...
nishat tahsin's user avatar
1 vote
2 answers
108 views

When to add signals coherently and incoherently?

I just learned in transmission electron microscopy about coherent and incoherent summation. I can't seem to wrap my head around the fact that intensities can be added incoherently. Why can you do that?...
Dat Ho's user avatar
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1 answer
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How to condition an STM tip?

I am starting work on an atomic force microscope and a scanning tunneling microscope and I wanted to ask for advice. How to proceed to get the sharpest tip and therefore the best resolution?
physics enthusiast's user avatar
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1 answer
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Limiting factors in traditional microscopy

I have been researching Entanglement-Enhanced Microscopy, and the overall breakthrough seems to be obtaining a resolution of $1/N$ as opposed to $1/\sqrt N$. This led me to ask what the limiting ...
murtadee hughes's user avatar
1 vote
2 answers
77 views

Image of an electron

Suppose we are trying to see the image of an electron with a microscope. Is it possible? And if it's possible then what do we see? A point at a time, or a blurry spot, or something else?
Users's user avatar
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Where is the final image located at in a compound microscope?

In a compound microscope, is there a standard as to where the final image should be located? After doing some research, I arrived at either the final image should be located at infinity (to be focused ...
Astrovis's user avatar
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What is the relation between image resolution and Cutoff from 2D FFT functions in frequency space?

This might be dumb, but unfortunately I need some urgent help about Cutoff from 2D FFT functions in frequency space. I am writing my bachelor thesis, near DDL and cannot get a lot of help offline in ...
Anothernewbie's user avatar
2 votes
2 answers
138 views

Why does Near Infrared (NIR) light allow deeper imaging in biological tissue in Multiphoton Microscopy?

I have been reading this paper on multiphoton microscopy (https://www.nature.com/articles/nphoton.an.2010.2) and I am very confused about something. Here is a quote from the paper: "The NIR ...
Sigma123's user avatar
3 votes
1 answer
70 views

Can a system of lenses or compound lens have a higher numerical aperture than a single element in the system?

I am attempting to create a simple Abbe condenser for a microscope, which typically consists of two converging lenses. Ideally, I would like to achieve a numerical aperture for the condenser of ...
Yashka Oreza's user avatar
1 vote
1 answer
66 views

Is reflection from pockel cell normal?

We are trying to align a laser for a 2-photon microscope and we noticed a reflection from the pockel cell. The reflection can be caught on the edge of the waveplate or the iris that are before the ...
Noushin Ahmadpour's user avatar
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3 answers
248 views

Imaging and illumination plane in microscope

I've been delving into the intricacies of achieving Kohler illumination in microscopy, and the term 'illumination and image-forming rays' consistently surfaces. Most resources emphasize the ...
CoffeeBiscuit's user avatar
3 votes
0 answers
107 views

Out-of-focus color in reflective microscopy with Köhler EPI-illumination

It is well known that Köhler illuminator in optical microscopy is designed in a way to ensure uniform illumination of the field, often with adjustable apertures to allow as little light as possible to ...
BarsMonster's user avatar
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Can the principle of helium ion microscopes be generalized to heavier noble gasses for suitable substances?

I have heard helium ion microscopes offer extremely high resolution of features due to the shorter debroglie wavelength of the helium atoms. Naturally I was curious if anyone has built something using ...
Sidharth Ghoshal's user avatar
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Beam diameter of a fluoresence emitter after an objective

I try to calculate the beam diameter after a high NA objective that focuses on a point-like emitter that emits fluoresence light. To my basic understanding, I get this by computing the Entrance pupil (...
P. Egli's user avatar
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Is it possible to make real DIY microscope with several lens?

I have alot of small double convex 7 mm lenses and i have very small lens 4mm And i have plano concave lenses all of them is small I tried to combining them to get a magnified image and it worked when ...
Pcp115's user avatar
  • 101
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2 answers
165 views

Trouble simulating/ray tracing a simple microscope

I'm trying to run a (seemingly) simple simulation of a basic microscope using the free software WinLens 3D Basic, but the ray tracing looks wrong: The microscope consists of a small plano-convex lens ...
srhslvmn's user avatar
  • 181
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1 answer
200 views

Why do fluorescence microscopes use long-pass dichroics

Superresolution fluorescence microscopy is very sensitive to chromatic aberration. Dichroic mirrors, while being biplanar, are not normal to the optical axis and so will cause some dispersion in one ...
selene flemming's user avatar
1 vote
0 answers
50 views

Anomalous Spectral Function and STM

I have a question in regard to the anomalous spectral function and how it pertains to STM experiments. The use of the spectral representation of Green functions is assumed known, so I do not go into ...
scruby's user avatar
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1 answer
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Question in Linear response theory

I am currently trying to do a calculation using linear response, but I am confused on one thing I never thought of. To start here is the relevant equation (I assumed the operators are self evident to ...
scruby's user avatar
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1 answer
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Near Field Optical Microscope

As a handyman, once I took pictures of a corner of a parfaim bottle by my camera (Near Field Optical Microscope), I'd like to know the interpretation of the following image: less powerful zoom: https:/...
The Tiler's user avatar
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3 votes
3 answers
854 views

Why can’t we see atoms in an optical microscope?

I know, the answer to this question may seem obvious: The resolution/magnification of an optical microscope is limited by the minimum wavelength one uses. This is due to the diffraction limit. However,...
Lockhart 's user avatar
1 vote
2 answers
155 views

Aperture/image circle in a DIY digital microscope?

I have just taken apart a broken USB microscope, and recognized that there is actually no magic going on inside. So I thought how about building one of my own. The idea is to buy a regular (possibly ...
oliver's user avatar
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1 answer
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(Optical microscopy) Resolution improvement by using oil immersion

'Oil-immersion' technique is widely used to increase the numerical aperture, and hence the resolution in optical microscopy. Some microscope manufactures like Nikon or ZEISS explain the principle of ...
quantumoptics666's user avatar
2 votes
0 answers
36 views

Can you use a digital camera CCD to make an image without a lens?

What would you be able to see if you took off the lens of a digital camera and placed a drop of pond water containing microbes/amoebas directly on the sensor? Would you be able to see an image? The ...
Rishi's user avatar
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0 answers
26 views

Sharpness of images on specular vs diffuse surfaces

I'm observing a confusing phenomenon where I'm projecting an image episcopically through an objective into a substrate while observing it through a camera (think structured illumination for a ...
lionelbrits's user avatar
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0 votes
1 answer
53 views

STED Microscopy: Differentiate between emission from stimulated emission and fluorescence

From what I understand about stimulated emission in STED microscopy, a red shifted Laser (apperently with a wavelength around the emission wavelength of the fluorophor) is used to induce stimulated ...
nebroth's user avatar
1 vote
2 answers
56 views

Laser confocal microscope filter

I wanted to mount a narrow bandpass filter which allows to transmit 405 nm wave, i. e. the laser wavelength. I wanted to cut off other light, cause my detector is also sensitive to other wavelengths. ...
user avatar
2 votes
1 answer
51 views

Does the substage condenser numerical aperture cap total resolution of an imaging system?

Amongst formulas to estimate the theoretical resolution of a transmitted light microscope, I often see: $$ \delta = \frac{1.22\lambda}{NA_{obj} + NA_{cond}} $$ Where $NA_{cond}$ is the numerical ...
ilykos's user avatar
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1 answer
152 views

How can I magnify an infinity corrected image onto a camera surface?

I know that the typical setup with a camera will place the CMOS sensor at I(3) in the image below, and that this magnification is dictated by the tube lens focal ...
joshp's user avatar
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0 answers
95 views

How do I read a scanning electron microscopy (SEM) image and an element of the photo

I'm trying to figure out the information I can take of this image. What I did found out is the following information : The increase is 600 times. The voltage is 30.1 kilowatts. The relief corresponds ...
Lea Katerina's user avatar
2 votes
3 answers
87 views

Operation of a microscope and image reconstruction

I have a question about the operation of the optical microscope. With reference to the image: from what I know, the image of the object is formed the first time where the red arrow is, why then is ...
Salmon's user avatar
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1 answer
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In a Mirau Interferometer what is the distance between the mirror and the beamsplitter?

A Mirau interferometer (shown left in the figure, next to the Michelson configuration to the right) is an important instrument for non-contact surface profilometry. Briefly, taken from Wikipedia, &...
user391339's user avatar
1 vote
0 answers
16 views

Is it possible to achieve Atomic resolution with the following parameters of a Scanning Probe Microscope? [closed]

I understand that the formula for Resolution is the Dimension of the image/Dimension of the object, however I am still unable to solve this. Thanks for any help in advance
ojiman's user avatar
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1 answer
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Formula for total magnification of a compound microscope

In the book 'Materials Characterization: Introduction to Microscopic and Spectroscopic Methods, Second Edition pdf' its says: The total magnification of a compound microscope as shown in Figure 1.1 ...
Jack Jack's user avatar
  • 187
1 vote
1 answer
70 views

How can can evanescent field cause fluorescence if there is no energy transport?

I am currently looking at the topic "Total Internal Reflection Microscopy". This creates an evanescent field travelling to my sample. This evanescent field no activates the Fluorophores. But ...
Tobias's user avatar
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1 answer
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How does coherence of a light affect how we perceive a sample in a microscope?

First of all, does coherence of a light, which is used for illumination in a microscope, affect the sample, or the way we see/perceive it? I want to know what optical elements do I need to use if I ...
Meera's user avatar
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1 answer
1k views

Why Airy disk (Fraunhofer diffraction) is first order of the Bessel function of the first kind?

I have been studying Fourier ptychography. I am confused about why Airy disk is J1 of the Bessel function. I think that Fraunhofer diffraction looks more like J0 of the Bessel function.
許煜承 Wistar's user avatar
2 votes
1 answer
402 views

How to set mask size when apply inverse-FFT to the power spectrum of HAADF-STEM image?

Hello everyone, I want to characterize dislocation in my FIB sample using Titan electron transmission microscopy and I want to use FFT filter to process my HAADF-STEM image with DigitalMicrograph (...
Joseph's user avatar
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