JPH065334B2 - Micro needle microscope auto focus device - Google Patents

Description

TECHNICAL FIELD The present invention relates to a microneedle microscope autofocus device mainly used in the field of biotechnology.

(B) Prior Art Conventionally, the focus position of the microscopic needle of the microscope has been manually operated while observing the display.

However, in such a focus adjustment method, in the field of biotechnology, which has been remarkably developed in recent years, a microneedle capable of invading cells can be efficiently focused while being held on the microscope stage. The alignment becomes extremely difficult, and therefore there is a disadvantage that the microneedle cannot be accurately introduced into a predetermined portion of the cell.

(C) Objective The present invention has been made in view of these circumstances.
One of its main purposes is the autofocus of a microneedle microscope, which can quickly detect the focus shift with respect to the tip of the microneedle placed under the microscope barrel and focus automatically. To provide a device.

(D) Configuration The present invention relates to a micro needle that is positioned below the lens barrel of a microscope and is held in a state of being inclined with respect to the stage, a micro needle moving means that moves the micro needle, and a micro needle tip portion near the video. Video pick-up means for picking up images and outputting video signals from a plurality of points is compared with rising times of the video signals outputted from a plurality of points, and a video signal having the shortest rising time is selected and calculated to select the selected video. The micro needle is moved in the direction of the optical axis of the signal, each of these calculations and the movement of the micro needle are repeated until the rise time falls within the preset allowable range, and the video imaging means and the micro needle moving means are operated. It is a microscope autofocus device for microneedles, which is provided with control means for instructing.

(E) Embodiments The present invention will be described in detail based on the embodiments shown in the drawings. The present invention is not limited to this.

FIG. 1 shows an embodiment of the present invention. A micro needle microscopic autofocus device (1) comprises a microscope (2), micro needles (3), video imaging means (4), and micro needle moving means. (5), and control means
It is mainly composed of (1) from (6).

The microneedle (3) is a holding member (9) between the objective lens (7) of the microscope (2) and the stage (8) so that the front end thereof is directed obliquely downward by an angle θ (for example, 30 °) from the horizontal direction. Installed on. The video imaging means (4) is a device for video shooting the tip of the micro needle (3), and is installed above the eyepiece (10) of the microscope (2). Video signals extracted and output from a plurality of points on the tip of the microneedle (3), for example, two points (A) and (B), as shown in FIG. 2, are input to the control means (6), where Fig. 3
The rising times (TA) and (TB) of the video signals for two points (A) and (B) as shown in (a) and (b) are compared. (V is voltage,
t is time). It is known that the rise time at the point where the video signal is in focus is shorter than that at the point where the video signal is out of focus. Therefore, in Fig. 3 (a) and (b)
If (TB) <(TA), it can be seen that the focal point at the tip is closer to (B) than (A). Therefore, the control means
If (6) is set to focus on the point (A) of the tip of the microneedle, in this case, the control means (6) moves the microneedle moving means (5).
A signal is issued to move the micro needle (3) upward together with the holding member (9).

On the other hand, when (TA) <(TB), it can be seen that the focus is closer to the (A) point than the (B) point, but the focus is between the (A) point and the (B) point. Since it is unknown whether it is outside (3), in this case, the control means (6) gives a signal to the microneedle moving means (5) to move it upward or downward, and about the point (A) after the movement. Depending on how the rise time of the video signal changes
The subsequent movement direction of (3) is determined. For example, if the rise time (TA1) after movement is longer than the time (TA) before movement, it is determined by the control means (6) that the movement direction of the microneedle (3) was opposite. A movement signal in the opposite direction is newly output to the micro needle moving means (5), and the focus position of the micro needle (3) is adjusted to the set point.

The operation command to move point (A) near the focal point is
This is repeated until the rising time (TAi) when the point (A) is moved i times satisfies the following equation.

(X−α) <TAi <(X + α) where (X) is the rise time when the focus of the video signal is obtained experimentally in advance, and α is the rise time indicating the allowable range of the focus. Is a permissible amount and is a value set in the control means (6).

At the point where the microneedle (3) is desired to be in focus, the control means (6) controls the microneedle moving means so that the rising time input from the video imaging means (4) to the control means (6) satisfies the above relational expression. By controlling (5), it is possible to automatically focus on a predetermined point of the minute needle (3). Thus, the position of the tip of the micro needle (3) can be accurately grasped, and the subsequent movement of the micro needle (3) can be performed accurately. In particular, the tip of the microneedle (3) can be accurately inserted into a predetermined location such as a cell.

In the above embodiment, there are two video signal extraction points for the microneedle (3), but if the number of extraction points is increased to three or more, the moving direction of the microneedle (3) for focusing is increased. Since it is easy to understand, the time required for focusing can be shortened.

If it is necessary to conduct research while observing the movement state of the micro needles (3), even if the display means (11) shown by the phantom line in FIG. 1 is connected to the video imaging means (4). Good.

(F) Effect The present invention extracts video signals from a plurality of locations near the tip of a microneedle held obliquely under the microscope barrel and determines the moving direction of the microneedle by comparing their rise times. Since focusing is performed, it is possible to automatically and efficiently focus on the tip of an elongated object such as a microneedle. Therefore, after such micro-needle focusing, that is, after initial setting of the micro-needles, if the cells are set to a predetermined position, the micro-needles can be accurately moved to the predetermined position immediately after the cells are focused. it can.

[Brief description of drawings]

FIG. 1 is a functional explanatory view showing an embodiment of the present invention, FIG. 2 is a front view of a main portion for extracting a video signal from two points of a microneedle tip, and FIGS. 3 (a) and 3 (b) are 3 is a graph showing rise times of video signals at two points (A) and (B) in FIG. 2. (1) …… Micro needle microscope auto focus device, (2) …… Microscope, (3) …… Micro needle, (4) …… Video imaging means, (5) …… Micro needle moving means, (6) ...... Control means, (8) …… Stage, (11) …… Display means, (θ) …… Inclination angle with the horizontal direction of the minute needle.

Claims (1)

[Claims] 1. A microneedle positioned below a lens barrel of a microscope and held in a state inclined with respect to a stage, a microneedle moving means for moving the microneedle, and a video image of the vicinity of the tip of the microneedle, The video image pickup means that outputs video signals from a plurality of points and the rising times of the video signals output from a plurality of points are compared and calculated, and the video signal with the smallest rise time is selected and calculated. Control for moving the micro needle in the axial direction, repeating each of these calculations and moving the micro needle until the rise time falls within a preset allowable range, and for instructing the video imaging means and the micro needle moving means to operate. And a microscope autofocusing device for microneedles.