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Phase contrast microscopy is used for observation of oral bacteria including periodontal disease bacteria.

Phase contrast microscope is a biological microscope that can observe by converting the phase difference of light into contrast.

You can observe specimens close to transparent bodies without dyeing.

 

By using a normal phase contrast microscope you can observe bacteria in the oral cavity.

 

There seems to be a company specialized especially for dental clinics especially among them.

http://www.ptech.jp/pscope/（web site in Japanese）

http://www.ipona.biz/dentist/dentist.html（web site in Japanese）

Metallic microscopes have more adjustment points than ordinary microscopes. Depending on the adjustment, the light intensity may decline extremely. If it is dark (or no light comes out) please check as the following instructions.

 

■Polarization observation change lever
When both levers protrude, the light path is blocked.
Be make sure to switch between polarization observation and normal observation.
■Optical axis adjustment of light source
	If the optical axis and the tilt of the light source deviate greatly, the light irradiated from the objective lens may be drastically declined.
■Adjusting the aperture in the middle of the optical path
	There are two aperture functions in the middle of the light path. If this is narrowed too much, the irradiated light may decay considerably.

 

It is possible by attaching option struts to the first arm of our Smooth Arm.

C clamp can be diverted to standard equipment. So, it can be mounted on the edge of table shelf.

An optional wall mounting bracket is also available.

C Clamp (standard equipment)	Wall mounting bracket (optional)

you can adjust the arm as above.

The microscope camera has a connector len at the top.

Therefore, you can attach to the eyepiece part of the microscope and the JIS lens.

connect to eye piece part	connect to Trinocular lens barrel (camera port)

 

You can also connect to the C mount by removing the connector lens at the tip.

Camera for the microscope consists of the three parts of the belows photo.

 

1. Camera body 2. C mount ring 3. Connector lens

Remove only the connector lens, be make sure to attach the C mount

 

In this picture, you can connect as a C  mount camera.

 

In this case, the lens is not attached to the camera. Therefore, you need to adjust the field of view on the C mount side (microscope side).

For details, please refer (“Connection of camera for microscope”).

When observing with a binocular stereomicroscope with both eyes, the objective lens is slightly oblique so that the object is close to the actual view.

(It is the same as looking at nearby objects with both eyes of a human.)

 

Therefore, looking at the following objects with one eye

 

 

You can see the side slightly. (In other words, you will be looking at diagonal.)

 

When the microscope is moved up and down, the field of view also shifts slightly.

The focus synthesis software is made on the premise that the field of view does not shift.
(It is assumed that it vertically moves up and down to the object)
If you use focus synthesis software with a stereomicroscope, it will blur like a photo below.

 

Mitani Shoji’s focal synthesis software (WinROOF 2018 Lite) has “microscope mode”.

Focus synthesis is possible without bleeding by focus synthesis using this function.

We also support these problems. Please feel free to contact us.

 

Of course, if you are a straight tube microscope like the one on the left, you can use normal focus synthesis software without problems.

Normally, focus adjustment of the microscope is performed when observe by naked eye.

Due to individual variations of the camera, the focal point of the naked eye observation may not match the focus of the camera.

In the case of JIS lens (see “Connecting camera for microscope”), the back focus is attached to the lens.
You can adjust easily.

 

In case of C mount, some of the microscope side C mount part has back focus, but it is very slight.

There is also a mechanism to adjust the back flange (back focus in this case) on the camera side. However, I think that this is also numerically small.

 

Eyepiece micrometer (reticle) is a glass scale used by incorporating it in an eyepiece when measuring the size of a specimen with a microscope.

There are various kinds depending on how to use it.

Corresponding size varies depending on the size of the microscope.

φ19, φ20, φ20.4, φ21, φ24, φ25, φ26, φ27 mm are general-purpose sizes and can be easily purchased with onlineshops.

For many eyepieces, you can insert later as needed.

Since there are some errors in the magnification of the objective lens, the measurement of eyepiece micrometer (reticle) becomes simple.

<How to use>

The width of the observed objects on a bar divides will be changed depend on the magnification of the objective lens.

Caculation:

Width on specimen of 1 scale (mm) = actual size of 1 scale of micrometer / magnification of objective lens

<Example>

Using a micrometer (0.1 mm / scale) of 10 mm / 100,

When a lattice sample of 0.2 mm pitch was measured with a microscope has objective magnification of 10 times

0.1 mm (actual scale of 1 scale) / 10 times (objective magnification) = 0.01 mm (1 scale on sample)

 

 < Measurement 10mm/100 (reticle) has step 0.1mm>

 

< Measurement 10mm/100 (reticle) has step 0.1mm>

 

Measure sample of 0.2 mm lattice

 

 

In 0.2 mm has 20 scales. (Precisely 19.5 scales)

The difference (about half scale) is due to the magnification error of the objective lens.

The camera port of the microscope is roughly divided into the following two types: JIS lens and C mount

(Besides that, there are other classic or foreign types)

JIS lens (straight lens diameter of 23.2mm)	C-mount (Screw M25.4mm)

If you want to connect a camera with a “C-mount” to the mount of a microscope, there are two methods:

Convert the camera port on the microscope side to C-mount:
If the microscope already has a C-mount, you can directly attach the camera to it.

However, if the microscope has a JIS (Japanese Industrial Standard) microscope tube, you will need to modify the port on the microscope side.

Please note that not all microscopes allow for port conversion, so it may not be possible in some cases.

 

 

 

 

The biggest advantage of this camera is to be connected to the eyepiece section.

 

 

 

 

<Camera for microscope attached to C mount>

C mount is one of mount camera standards. (Please refer to “C mount, CS mount”)

Because of mount camera standards, a microscope camera is unnecessary.

It is the greatest merit that C mount type general purpose camera can be connected as it is. (Can not be connected to eyepiece section.)

 

 

However, the field of view (magnification) will change depending on the size of the image sensor on the camera.

As there is no lens on the camera, the magnification can not be adjusted.

(It must be selected when purchasing the camera adapter for microscope)

The following picture is a system diagram of some microscope manufacturers.

C-mount has various types from 0.25X to 0.63X.

It is necessary to purchase C mount matches to the number of inches of C mount camera.

Since it’s all C mount, the focus themselves matches. However, the field of view will be narrow or vignetting (shadows of four corners of the image) occurs.

 

2.  Make a C-mount camera compatible with a JIS (Japanese Industrial Standard) microscope tube

When you attach a JIS microscope tube adapter lens to your camera, you can mount it onto the JIS microscope tube.

The biggest advantage of this camera is that it can also be connected to the eyepiece lens section.

If you remove the front lens, you can also connect it to a C-mount. Please refer to the instructions on “How to Connect a Microscope Camera to a C-mount” for reference.

3. Field of View Considerations:

 

Camera internal sensors (CMOS or CCD) come in various sizes. Ideally, the lens magnification should be adjusted according to the sensor’s size.
If the sensor size is 1/2 inch, it is recommended to choose a lens with approximately 0.5x magnification.

 

 

If you have other inch sizes or specific requirements, please contact our technical support.

 

High definition video will be horizontally image of 16: 9. (1920 X 1080)

 

The field of view of the stereo microscope is a round view.

 

To take pictures of a microscope with a Full HD camera, there are two following methods.

1. If shooting all fields of view of the microscope, put out small images on the monitor. (The resolution will be about ¼)		2. If shooting to avoid dark conrner, It will be smaller than the actual field of view. (In fact, it is enlarger) it will be smaller than the field of view
Field of view observe with the naked eyes		HDCE-20T Full HD camera images for the HDCE-20T microscope

 

If you want to extend the field of view, you should use a 4: 3 camera (PC monitor direct with (analog RGB) camera or video camera) and se a len with a wide field of vision. (Some vignette will come out.)

 

Below picture is microscope camera with X 0.36 lens attached to our PC monitor direct (analog RGB) camera.

* Shadows appear in the four corners. Resolution is also lower than Full HD.

 

Use a 4: 3 camera and a wide field of vision len, the image likes the picture below.

I will explain the field of view of a camera of microscope attached to an eyepiece lens or JIS lens.

(For camera of microscope which C mount is attached, please refer to “Connection of microscope and C mount camera”.

 

The left picture shows the image using microscope with the naked eye.

1. Camera of microscope
Normally, cameras for microscope are made in order to avoid vignettes (shadows of the four corners) It is the red frame of the top. Although it designs with margin in order to avoid vignetting, its level of variation depending on the manufacturer. However, it is enlarged more reliably than the naked eye observation. Because the magnification of the lens of camera for microscope is fixed, it is not possible to change the field of view.
2. Commercial video camera and connector lens
I am observing it by installing a connector len on a commercially video camera. Commercially video cameras have a zoom function. Using this zoom function, we adjusted the field of view. Vignetting (shadow of four corners) occurs, but you can take a wide field of view. (Yellow frame of the top photo) You can export output signal to the monitor using the HDMI cable from the video camera.
HDMI Cable

Firstly, please check the proper eye position. This eye position is called eye point.

With the old microscope, the eyelashes were close to touch the eyepiece. Now, there are various high eye point that you can observe while wearing glasses.

 

If the eyepiece contains the following marks, it is a high eye point.

 

I will show you one way to easily know eye points.

Place copy paper on the eyepiece.

Changing the distance of the copy paper from the eyepiece changes like the photo below.

The point with the most focus is the eye point.

< Focus status>	< Unfocus status>

Place your eyes at the correct position (eye point) then adjust.

１．Inter-eye adjustment (Adjust the eyepiece and eye width.)

When looking at the binocular microscope, there is one tip is to look far away.

Looking into the eyepiece while consciously looking far away then adjust the width of both eyepiece lenses.

 

I looked through the eyepiece with both eyes and the adjustment is complete once the field of view becomes one full circle.

２．Diopter adjustment

Adjust by turning the diopter adjusting ring so that both eyes are in focus.

(One-sided diop adjustment)	(Two-sided diop adjustment)

３．Zoom adjustment

(1) Turn the zoom dial to set the minimum magnification and focus.

(2) Next, turn the zoom dial to set the maximum magnification and focus.

(3) Return to low magnification again. Adjustment is completed if the focus is on the low magnification side.

When the focus is out of focus

Repeat steps (1) and (2) for one-sided diop adjustment.

If it is a two-sided diop adjustment, turn the diopter adjusting ring so that the two eyes are in focus at the stage (3).

Then repeat the operations from step (1) to (3).

The magnification of the optical microscope and the magnification of digital microscope are very different.

The magnification of optical microscope is absolute magnification, even if the manufacturer differs, if the magnification is the same, it will be in the same field of view. (It is an absolute magnification.)

However, for digital microscope, the field of view will be very different depending on the manufacturer even at the same magnification. (Relative magnification.)

■Optical Microscope magnification

The magnification of the microscope is calculated by the following formula.

Optical magnification = (magnification of the objective lens) × (magnification of the eyepiece) Magnification of the microscope = optical magnification.

(In the case of the microscope of the bottom photograph, the magnification becomes 10 times the magnification of the objective lens × the magnification of the eyepiece 10 times, the optical magnification becomes 100 times.)

 

Even in the case of a microscope, when the camera is attached to microscope, it may be expressed including the monitor magnification.

(Refer to “How to calculate magnification of camera for microscope”.)

■Digital Microscope magnification

Taking an example of a microscope with an objective lens, which is easy to understand.

Firstly, the calculation of optical magnification is the same as the microscope.

Optical magnification = (magnification of the objective lens) × (magnification of the lens)

(In the microscope of the below picture, the magnification of the objective lens is 10 × magnification × 2 times the magnification of the lens body, the optical magnification is “20 times.”)

 

However, the monitor magnification is shown here.

Overall magnification = monitor magnification × optical magnification.

If the image sensor size is 1/2 inch and the monitor is 17 inches, the monitor magnification is 54.

(Please refer to “magnification of microscope”.)

Overall magnification = monitor magnification × optical magnification = 54 × 20 = 1080 times.

In conclusion, if the monitor size or image sensor size changes, the magnification will also change.

(Looking at the catalog of each manufacturer, there is a note such as “Equivalent to” ○○ inches monitor “

< Magnification definition>

Computing is complex, but the way of thinking is simple.

Microscope as shown below with 1mm on the display to 40mm.

This is simply 40 times.

 

 

Summary: 

 

The magnification concepts of a microscope and a macro lens are different and not interchangeable.

 

The magnification of a microscope is calculated as: 

Optical Magnification = (Objective Lens Magnification) × (Eyepiece Lens Magnification).

 

The magnification of a macro lens is calculated as:

Optical Magnification = (Objective Lens Magnification) × (Lens Body Magnification).

 

In the case of a microscope, the overall magnification includes the consideration of monitor magnification in addition to the optical magnification.

 

Total Magnification = Monitor Magnification × Optical Magnification.

 

■Adjust the light source position
(BEFORE ADJUST)	(AFTER ADJUST)
■ Adjust lens position of the light source
(BEFORE ADJUST)	(AFTER ADJUST)
■ The image sensor size of camera Even the image sensor size of camera changes. Reducing the image sensor size, the influence of variations in illuminance distribution may be reduced. However, its picture will be enlarged than the actual naked eye observation.
Camera sensor 1/2 inch	Camera sensor 1/3 inch
(Note) Since the cameras are different, color and contrast are slightly different.

 

With normal microscope, the distance from the lens center to microscope stand is limited. Thus, if it is a wide object, it will hit the stand.

 

At our company, we have options (special order) like the theone on the right picture.
This option can be installed both vertically and horizontally on our large stand.
■Set up on the long side
note: When using in this direction we recommend attaching a weight (optional) to stabilize
■Set up on the short side

 

Considering the filter that cuts the wavelength of illumination (470 nm, 505 nm) and allows only the emission wavelength to pass. It will be below.

SC 56 is Fuji Film’s model number.

The equivalent one is model O56 of HOYA company.

We can install it to our stereo microscope too.

It is not specified as GFP, but the characteristics are equivalent.

We will show you how to attach translucent light to the base without transillumination afterwards.

(There are some conditions based on base shape.)

As mentioned above, there are the following methods as long as it is a microscope with an observation board (black and white plate).

Firstly, remove the observation board.

Place the surface emitting light under the base. Pull out the cable from the gap.

Attach the glass plate instead of the observation board.

Adding a glass plate and surface emission lighting can be used as a simple transparent lighting stand.

 

 

<Transmitted lighting used>

Light illumination for small camera stand RD-95T

 

<Stand type>

If it is a stand type, you can turn it to the side if you attach (“3D arm”) in order to replace with a stand of a wide base with good stability.

 

<Arm type>

If you use our smooth arm, you can easily turn it to the side.

The RMS (Royal Microscopical Society) standard is the specification of the objective lens connection part (M20.32 mm P = 0.7.6 mm) of the microscope.

 

RMS → C mount

RMS → T mount

 

Conversion adapters such as are commercially available.

You can do various things by using this.

There are scales on the software.

The pitch of the scale can be changed

The above picture is adjusted to divide the 0.1 mm width into 25 equal parts. (You can not put numbers on the scale.)

 

（１）Shooting through a reticle (glass scale) lens with a small digital camera

 

There is also a way to use ordinary digital camera and camera bracket.

 

Using this bracket, and attach a digital camera to the eyepiece of the microscope and shoot.

 

 

 

 

(2) Shooting with a microscope camera that covers the eyepiece

 

Place a eyepiece (glass scale) of the microscope.

Cover the eyepiece with a microscope camera that covers the eyepiece.

 

 

Then you can take pictures with a reticle (glass scale)

■Focusing

Focusing during microscope shooting is not a microscope eyepiece but please go while watching the camera’s LCD screen. (It will be a viewfinder on cameras without live view function.)

*auto focus function of camera doesn’t work at this time. so please adjust a focus with using focus angle of stereo microscope.

 

Even if you focus on an eyepiece, it usually does not match the focus of the camera.

(If there is diopter adjustment or back focus on the microscope, tonality may be possible.)

■Vibration reduction

It will be like a picture blurred when you shake hands.

To prevent camera vibration, please use remote control or set the time settings.

For cameras with low vibration mode, set the low vibration mode to ON (about 1 second).

■Exposure adjustment

The exposure priority mode (dial A) is easy when shooting. If the monitor is too dark or too bright, please adjust the light / dark control (±).

Depending on the camera, “lens unmounted” may become occasionally.

In this case, shoot in manual mode.

(In this case, the brightness is adjusted by the shutter speed.)

■White balance

Please manually adjust if the image becomes reddish or blue-colored due to the color of the light source of the microscope.

Make the light source of the microscope proper, remove the specimen, set it to nothing on the screen (glass part without specimen in the case of a biological microscope, white or gray board in case of a stereomicroscope), set the screen to white I will let you recognize.

※ Please see the operation manual of the camera for detailed operation.

■ISO sensitivity

If the ISO sensitivity is too high, the image quality will degrade. If the screen is dark, you can take a beautiful image by using some method such as brightening the light of the microscope or slowing down the shutter speed, setting the sensitivity to a lower setting and shooting.

When you observe the microscope with the naked eye, the field of view gets rounded.

(We are using glass scales in increments of 0.5 mm as test samples.)

 

 

 

When this is shot with a camera, it becomes a 4: 3 rectangle with enlarger size.

Besides that, the field of view will change depend on the magnification of the camera connection (connector lens), the size of the image sensor of the camera, and so on. (It will also be a red field of view and a blue field of view in combination.)

You can express this with monitor magnification. (You can also calculate.)

■Camera connection part is C-mount

If the camera connection part on the microscope is C mount, the C mount part also has magnification.

If you do not specify microscope, it seems that a C mount adapter of equal magnification (1 times) is often attached.

Monitor magnification = magnification of objective lens × magnification of C mount × (diagonal length of monitor / diagonal length of camera sensor angel)

 

1 / 2.5 inch camera (diagonal length of camera sensor angel is 7 mm)

19 inch monitor (diagonal angel 470 mm),

C mount part (equal magnification),

Objective lens 10 times

<Magnification caculation>　

10 times X 1 times X (470 mm / 7 mm) = 670 times

The field of view will be like the bottom picture.

 

 

Since the horizontal dimension of the 19 inch monitor is 370 mm. So, the image on the right spreads to the screen

<Measured magnification>

370 / 0.55 mm = 672 times

 

Magnification caculation (monitor magnification) and actual magnification are the same.

We need to reduce the magnification of C-mount in order to increase the field of view

We also offer equal size (standard equipment) and 0.5 times (option).

There are various magnifications when looking at other company’s catalogs.

■If you are connected to microscope by JIS len, the magnification can not be changed (equal magnification)

If you are connected a camera to this lens barrel, a relay lens is attached to the camera side. There is magnification in this relay lens.

 

Magnification calculation is as follows:

Monitor magnification = magnification of the objective lens × magnification of the connector lens × (diagonal length of monitor / diagonal length of camera sensor angle)

 

1 / 2.5 inch camera (image sensor diagonal 7 mm),

19 inch monitor (diagonal 470 mm)

Connector lens (0.45 times),

Objective lens 10 times

 

<Calculation magnification>

10 times X 0.45 times X (470 mm / 7 mm) = 302 times

Since the horizontal dimension of the 19 inch monitor is 370 mm. So, the picture on the right spreads to the screen then:

<Measured magnification>

370 / 1.2 mm = 308 times

Magnification caculation (based on magnification on monitor) and actual magnification are the same.

Our company has 0.45 times and 0.37 times connector lens available.

We’ll advise the suitable one to the customer according to the size of the image sensor of the camera.

You can see the visual field of the naked eye observation of the microscope is the right picture
Normally, a 1/2 inch camera and a 0.45-megapixel lens will be included in the kit. In this case, the field of view will be as shown on the right. With a connector len of 0.45 times, vignetting (shadow) does not occur at the four corners of the image.
In case you really want to get a wide field of view, if you attach a wide 0.37x relay lens (wide angle) to the 1/2-inch camera, it will look like the right picture. * Slightly, ambient distortion occurs.

For monocular len microscope, there are indirect lighting devices such as dome lighting and arch lighting.

However, for a binocular stereomicroscope, it is structurally two light path designs.

Therefore, the lighting device like the one above can not be used as it is.

There are also a way to weaken the reflection by using the LED (flat type) with the LED pointing straight down as below.

For simplicity, there is also a method to observe by attaching a diffusion tube to the LED ring light

When attaching general-purpose LED ring light (not special LED ring light) to the tip of a stereo microscope, if the inner diameter of LED ring light is larger than the tip part of the microscope (the red circle part of the below picture), basically it can attach.

 

(Microscope with attachment part at the tip)

 

Based on the type ofmicroscope, there is a T-bar hanging at the head or the head is larger than the inner diameter of the LED ring light.

 

(Microscope without attachment part at tip)

In such a case, you can easily attach the lighting by attaching the LED fixing ring using the inner screw of the microscope.

For microscope as described above, the inner screw for fixing the filter is cut on the back of the tip cover.

As it is for optical filters, pitch microscope is common regardless of microscope manufacturer. (P = 0.75 mm)

* By the way, it is a fine pitch than fine thread of he general ISO screw.

 

This can be used to attach the fixing ring.

(We have a fixed ring of M49 and M48 available.)

 

It is easy to attach LED ring light.

 

 

The inner thread diameter may be large with a very large microscope.

There is no problem in this case.

Step-down rings are on sale from filter makers and others.

 

The following example is a step down ring from M 58 to M 49 mm.

 

 

By combining an LED fixing ring (attach with step down ring M49), you can easily fix the LED ring light to a microscope with a large aperture.