Are you thinking that your desired protein is working well? But one cannot get it due to the limitation of the resolution of one’s confocal microscope. If you want to see a clear view, you have to go for the STED microscopy power. It is far better than a confocal microscopy power.
However, STED ensures a high resolution to get a clear image. Now you may want to know the working process of this STED microscopy. To know that you have to stay with us till the end of this content. So, let’s begin the discussion before you look for phase plates.
Working Process of STED
Before you go for the STED, you need to know the working process of it. It is very vital to get a clear concept about anything. First of all, we want to that the STED microscopy made from the source of a laser-scanning confocal microscope.
Now we have to know about such a kind of confocal microscopy. So, keep your eyes below! Usually, a laser-scanning confocal microscope works when the object focuses on the laser light.
And, you get the laser light against one small part of the sample. Even the intensity of the light distributes as stated by the PSF and the image resolution limitation.
After that, the produced fluorescence collected from the object and then referred to the sensor. Mainly, it refers to the single pixel. However, to get more pixels, you have to move the focus spot position.
Different Kinds of STED Microscopy Laser Beam
There are different types of STED laser beam available. So, in the below, we will try to introduce all the laser beam to you. Here, we will discuss three STED laser beams, such as pulsed STED, CW STED, and GCW STED.
Pulsed STED
Firstly, we will talk about the pulsed STED laser beam. Mainly, STED makes by using the pulsed beam lasers. This kind of laser is very standard and can repeatedly work to emit pulses from a similar duration.
That means it ensures a high resolution to show the view. Also, the two laser beam become optimistic and then hit the target. Then it works for the same duration that helps to make the high resolution.
Continuous Wave STED
Now let’s talk about the CW STED laser beam. This kind of STED can stay constant for a long time. The Leica implemented such kind of STED commercially. Usually, continuous-wave STED does not require preparation and optimization like the pulse STED.
The CW STED’s downside has no time to get enough photons before it starts to produce fluorescence. As a result, the shape of the doughnut comes into the focal point. That causes one slight blurring to get the last images.
Gated Continuous Wave STED
Lastly, we will discuss the GCW STED. First of all, it works in one way, and in some cases, it may become a problem. It follows time-gated discovery to record fluorescence.
That means one has to tell the sensor not to gather photons. Instead, it allows one to omit the rest of the photons that acted before the laser beam (STED) has time for taking full effect.
