ES2278483B2 - Morphometry system for semi-automatic acquisition of three-dimensional coordinates, has semi-rigid sensor and optoelectronic sensor - Google Patents

Abstract

The morphometry system has a semi-rigid sensor and an optoelectronic sensor. The semi-rigid translation broadcast is produced by rotating the micrometer screws of the microscope plate. The optoelectronic sensor quantifies the displacement of the approach. The output of the sensors is transmitted to the control console connected to a computer. The computer program communicates with the console to a text file with the coordinates selected in the workshop. The origin of coordinates is specified at the beginning of work by assigning zero to the sensors.

Description

Acquisition morphometry system semi-automatic three-dimensional coordinates.

Object of the invention

Acquisition morphometry system semi-automatic three-dimensional coordinates.

It is a system for selection semi-automatic structure three-dimensional coordinates microscopic in optical microscopy.

The object of the invention is the extraction of Simplified models of microscopic structures for analysis in a simple and economical way. The system can be coupled to different microscopes and their use is very intuitive for the Username.

The invention is applicable in the field of Anatomy and Histology.

Background of the invention

In most quantitative analyzes of Real images in optical microscopy is essential to simplify the information contained therein. One of the most methods used is to obtain the Cartesian coordinates of the significant points of the image. These coordinates are acquired usually manually, although there are also systems of automatic and semi-automatic acquisition. Starting of these coordinates, information can be obtained not only from the spatial distribution of the sample of interest, but also of other spatial parameters such as distance between points, areas delimited by polylines, centers of gravity, etc.

In Neuroanatomy, the pioneer in this type of analysis was Bok (Bok, ST, 1936, The branching of dendrites in the cerebral cortex. Proc. Kon. Ned. Akad. Wetenschap 39: 1209-218), who manually recorded the coordinates ( x, y). Glaser and Van der Loos (Glaser, EM, Van der Loos, HA, 1965, A semi-automatic computer microscope for the analysis of neuronal morphology. IEEE Trans. Biomed. Eng . 12: 22-31) were the pioneers in obtaining the coordinates automatically, directly from microscopic preparations, adapting to the microscope transducers that measured the linear displacement of the plate in the three axes of space. Garvey et al. (Garvey, LS, Young, J., Simon, W., Coleman, PD, 1973, Automated three-dimensional dendrite tracking system. Electroencephal. Clin. Neurophysiol . 35: 199-204) and Wann et al. (Wann , DF, Woolsey, TA, Dierker, ML, Cowan, WM, 1973, An on-line computer system for the semi-automatic analysis of Golgi-impregnated neurons. IEEE Trans. Biomed. Eng . 20: 233-247) coupled engines step by step controlled directly by a computer, which meant an important advance in automation since the movement of the microscope plate could be controlled from the computer itself. Subsequently, Overdijk et al. (Overdijk, J., Uylings, HBM, Kuypers, K., Kamstra, AM, 1978, An economical, semi-automatic system for measuring cellular tree structures in three dimensions, with special emphasis on Golgi-impregnated neurons J. Microsc . 114: 271-284) interposed a controller between the computer and the motors, so that the computer no longer needed exclusive dedication to move the plate. Since then, the trend in laboratories has been to use motorized plates. However, given the cost of motorized plates, the semi-automatic system remains the most widespread. In this type of system, the displacement of the plate and the focus of the preparation is done manually and the coordinates (x, y) are read directly on the image turned on the computer screen with the help of a mouse and the Z coordinate using a transducer connected to the micrometer screw of the microscope.

Description of the invention

The semi-automatic morphometry system allows the acquisition of models of microscopic structures three-dimensional This system is used in the industry biotechnology in situations where it is not possible to use of an automatic image analysis system. This invention is the reduction of a semi-automatic system of morphometry, eliminating the continuous connection with the computer, working on the microscope and recording the movements in the three directions of space with three sensors whose coordinates are stored in a buffer. The advantage of this way of working is that the system uses its own image microscope, eliminating spatial resolution problems that they impose cameras and graphics cards when working on the computer when the microscope image is transferred to the screen. The invention is designed so that it can be used on different microscopes and the information of each work session it is stored in the buffer of the control console that transfer the data to the computer through the serial port of communications Temporary disconnection of the control console of the electric fluid does not cause data loss. Optionally it is possible to work with a connected computer and, in this case, the Data transfer is done while working with the microscope

The system can work with one, two, or Three sensors at once. Semi-rigid sensors for linear displacement in X and Y adhere in one of their ends to the fixed base of the microscope plate while The other end of each sensor adheres to the mobile support. With this arrangement, any displacement of the plate will produce an elongation equivalent to the displacement in the sensors. For the rotation produced by the microscope focus screw a stepper motor is usually used, although it is also possible to use a shaft with flexible connection for a counter Opto-electronic rotational.

Description of the figures

Figure 1 shows the general layout with all the elements for the semi-automatic acquisition of three-dimensional coordinates with the invention.

Figure 2 shows the location detail of the semi-rigid sensors that detect the linear displacements X and Y of the plate and bushing coupled to the micrometric screw of the microscope focus.

Figure 3 shows the external appearance of the control console

Figure 4A shows a projection in opto-electronic sensor perspective (Figure 1, item 13) used for offset detection produced in the microscope approach. Figure 4B shows the side view (right panel) and front view (left panel) of the opto-electronic sensor.

Figure 5A shows a projection in perspective of the internal arrangement of the sensors semi-rigid (Figure 2, elements 11 and 12). The Figure 5B details the front view (left panel) and the view side (right panel) of the sensors semi-rigid

Figures 6A, B, C, D and E show the scheme General electronic circuit of the control console (Figure 1, item 2).

Figure 7 shows the connection scheme of the microprocessor (U1, Figure 6A).

Figure 8 shows the connection scheme of the control console memory (Figure 1, item 2).

Figure 9 shows the connection scheme of the screen (Figure 3, item 21; LCD1, Figure 6A) of the console control (Figure 1, item 2).

Figure 10 details the component connection responsible for the transmission of data via RS232 to the computer (U3, Figure 6E).

Preferred Embodiment of the Invention

The working arrangement of the invention is shown in Figure 1. Attached to the microscope (Figure 1, element 1) are the two semi-rigid sensors (Figure 1, elements 11 and 12) that detect lateral displacements in the X-axis (Figure 1, element 11) and Y (Figure 1, element 12) of the plate (Figure 1, element 15) of the microscope. The Z-offset detection is performed by the opto-electronic sensor (Figure 1, item 13) that is attached to the micrometric approach screw with a socket (Figure 1, item 14). The information of the linear displacement of the plate and the rotational information of the focus are transmitted to the control console (Figure 1, element 2) where it is processed and stored temporarily according to the selection of the type and graphic attributes preset by the user. The coordinates are stored in the control console memory (Figure 1, item 2) when the Pedal key is pressed (Figure 3, item 29) or when the pedal is operated (Figure 1, item 3), which facilitates the work on the microscope when freeing your hands. The control console (Figure 1, item 2) can optionally be connected to a computer (Figure 1, item 4) to transfer data without temporary storage. Figure 3 shows the detail of the external aspect of the control console, which is formed by a plastic and aluminum case with a reduced keypad and a liquid crystal display of 32 characters (Figure 3, item 21). In the upper left corner there are two LEDs, one red with the legend Power (Figure 3, item 22) that indicates when the device is connected to the electric fluid, and another green with the legend Data (Figure 3, item 23) that indicates when data is transferred from or to the computer. Below the screen, there is the keyboard formed by five keys to select the work modes: the first, Menu (Figure 3, item 24) selects the operation menu, the second (Figure 3, item 25) and the third ( Figure 3, item 26), with up and down arrow symbols respectively, are cursors for scrolling through the options presented on the screen. The fourth key, Cancel (Figure 3, item 27) is used to override an entered value, or exit a menu option. And the fifth key, Accept (Figure 3, item 28) is used to confirm the information that appears on the screen. Finally, the Pedal button (Figure 3, item 29) replicates the coordinate input function
current that the pedal performs (Figure 1, item 3).

Figure 4A shows a projection in opto-electronic sensor perspective (Figure 1, item 13) used for offset detection produced in the microscope approach. In Figure 4B you have the side view (right panel) and front view (left panel) of the opto-electronic sensor (Figure 1, element 13). The micrometric screw of the microscope approach is coupled by means of a bushing (Figure 1, item 14) that is welded to a shaft (Figure 4, element 131) with flexible connection, allowing absorb the height difference between the micrometric screw of the microscope and counter focus opto-electronic The rotation made in the Microscope approach is transferred to a grooved disk (Figure 4, element 133) that transforms the turn into pulses light-darkness that are detected by the photosensor (Figure 4, item 132) and sent to the counter opto-electronic (Figure 4, element 134) that Quantify the number of steps and the direction of the turn.

Figure 5A shows a projection in perspective of the internal arrangement of the sensors semi-rigid (Figures 1 and 2, elements 11 and 12). The  internal (Figure 5, element 113) and external cylinders (Figure 5, element 114) are represented in this figure in a wire model for easy viewing. The zero position of the sensor is specifies through the system management program (usually, in the middle part) so that the maximum distance traveled in the displacements in both directions are the same. Extreme distances occur when the inner cylinder (Figure 5, item 113) is completely inside the cylinder outside (Figure 5, item 114) and when the inner cylinder (Figure 5, element 113) is completely visible because it is totally out of the outer cylinder (Figure 5, item 114). Both cylinders (Figure 5, elements 113 and 114) are assembled to two clamping structures (Figure 5, item 111) that will be the points of attachment to the plate and the microscope. This clamp is performs with the help of adhesive sailboat tape on the underside of the clamping structures (Figure 5, element 111). The microscope plate shifts become equivalent displacements of the position cursor (Figure 5, element 112) of the potentiometer (Figure 5B, element 115), and so therefore, in variations of the signal transmitted by the potentiometer (Figure 5B, element 115). The view is detailed in Figure 5B front (left panel) and side view (right panel) of the linear displacement sensor, as well as the location of the potentiometer (Figure 5B, item 115) and its position cursor (Figure 5B, element 112).

The electronic circuit that governs the operation of the invention, is housed within the control console and has been mounted on a circuit board 160 x 100 mm double-sided print drilled with 0.8 bits mm, 1 mm and 1.2 mm in diameter. Sockets for fixing the integrated circuits and the welding of the components on the Printed circuit has been made with tin.

The main component of the electronic circuit is the microcontroller of the Microchip brand, model PIC16F876 (U1, Figure 6A and Figure 7). This device has 8 Kbytes of program memory and 256 bytes of RAM (Random Access Memory) for the variables of the management program of the invention. The 16 digital input-output lines of the U1 microcontroller are programmable in two 8-bit ports (RB0 to RB7 and RC0 to RC7, Figure 7). The 16 digital inputs are used for the treatment of keyboard signals (SW1 to SW8, Figure 6D). Of these 8 inputs are used in this invention since SW1 to SW5, leaving SW6, SW7 and SW8 for possible extensions. In the Figure 6D details the connection scheme of these inputs. TO digital input RC0-2 (Figure 7) connects the information transfer diode (Figure 3, item 23; D5, Figure 9). The power diode (Figure 3, item 22; D4 Figure 9) is connected to the power supply output (REG1, pin 3, Figure 6C). Finally the sensor opto-electronic (Figures 1 and 2, item 13) are connect to multiplexed outputs RB0-0 and RB0-7 (Figure 6D). The analog inputs of the microcontroller (RA0-0 to RA0-5, Figure 7) are enabled for the converter 10-bit internal analog-digital that allows directly read the absolute value of the position of the sensors semi-rigid (Figures 1 and 2, elements 11 and 12). He semi-rigid sensor for linear displacement in X (Figure 2, item 11) is connected to RA0-0 and the semi-rigid sensor for linear displacement in Y (Figure 2, item 12) is connected to RA0-1. In addition, the analog inputs (RA0-0 to RA0-5, Figure 7) are multiplexed as outputs digital to manage the liquid quartz screen (LCD1, Figure 6C). Conversion time Minimum analog-digital is 20 µS. The Microcontroller power (REG1, Figure 6C) is 5.25v maximum and 3.3 v minimum.

In the microcontroller program memory is the code that manages the inputs and outputs and the device operation. The device has 256 bytes permanent memory where the read values of each sensor, current working mode and configuration options user defined.

The selected data during a session of work when a sample is analyzed they are stored in a memory (U4, U5, U6, U7, Figure 8) type 24LC256 flash type of the 32 Kb Microchip brand, expandable another 128 Kb.

On a liquid quartz crystal display (LCD1, Figures 6A and 9) of 32 characters (Figure 3, item 21) are View system information. This screen is lit from behind, so that the information is visible in any lighting condition This screen shows the information concerning the operation and selection of the three-dimensional coordinate that is detected by the position of the sensors

Communication with the computer is done using the MAX 232 device (U3, Figures 6E and 10) which is a MAXIM brand chip, which converts 5v TTL signals to levels +12v and -12v according to standard features RS-232, for communication with the computer. This device establishes communication bi-directional between the control console (Figure 1, item 2) and the computer, through the DB9 connector (P1, Figure 6E) for transferring data from a session.

The console keyboard consists of five keys (SW1 to SW5, Figure 6D) that are used for the selection of The menu functions. The method of reading the pressed keys it is done by means of a sweep that the system performs sequential row check and subsequent column reading of so that when a key is pressed, the microcontroller (U1, Figures 6A and 7) detects the event and acts on the internal program to Perform the desired operation.

The voltage regulation circuit used device 7805 (REG1, Figure 6C) provides 5 v stabilized from a variable power supply between 7v and 14v of continuous voltage and it is the one that sets the supply voltage for the control console (Figures 1 and 2, item 2) and the three sensors (Figures 1 and 2, elements 11, 12, 13). The implementation of the electronic circuit of the control console (Figure 1, item 2) is performed on a fiber printed circuit board 1.2 mm thick glass and 100 x 160 mm dimension.

Finally, several connectors are available in the control console housing for communication with the external elements of the invention: the selection pedal (Figure 1, element 3), the semi-rigid sensors (Figure 2, elements 11 , 12), the opto-electronic sensor (Figure 2, item 13), the computer (Figure 1, item 4) and some auxiliary connectors for device checks during operation. The pedal (Figure 1, element 3) is connected by an RCA connector (J3, Figure 6A), the sensors (Figures 1 and 2, elements 11, 12, 13) by three connectors (J3, Figure 6A) type " Jack " 3.5 mm female and 3 terminals. The power supply from an external voltage reducer with transformer is connected to a coaxial connector (J1, Figure 6C) of
3.5 mm

The program stored in the microcontroller is has implemented in PIC BASIC language of the CCS firm.

Description of the mode of operation with the invention

The operator of the invention uses a marker inserted into one of the microscope eyepieces. This item allows to observe a pointer on the microscopic preparation. TO difference in the use of the cursor that is made in the programs graphics, in this invention, the pointer is fixed and what moves it is the image of the preparation, so that the operator uses the micrometric screws of the microscope to displace the preparation and place the pointer in the place where it Find the structures you must select.

At the beginning of each work session, the first operation to be performed is the definition of the origin of the coordinate system and the objective with which it works. To do this, first select the INITIALIZE option in the control console menu, which assigns the zero value to the counters of the three sensors and then select the OBJECTIVE option, which allows the selection of the calibration of one of the objectives available in the microscope. Next, the operator selects the type of items to be entered, choosing the CLASS option from the menu. The available possibilities are three: A (Open Polyline), C (Closed Polyline), P (Point). As alternative classification parameters, two numbers from 1 to 10 can be chosen. Once the working mode is established, the operator moves the preparation using the micrometer screws of the microscope and for each element that he wishes to store he will press the pedal (Figure 1, element 3 ) or the PEDAL button (Figure 3, item 29) on the control console (Figure 1, item 2). The selection operation involves the storage of the absolute positions of the semi-rigid sensors (Figure 1, elements 11 and 12) and the relative position of the opto-electronic sensor (Figure 1, element 13) of the focus. At the end of the work session, the operator can keep the session data stored in the control console memory, or transfer the data from the control console (Figure 1, item 2) to the computer (Figure 1, item 4) through the serial communications port.

For the transfer of the data to the computer, the user will select the SEND menu option that prepares the protocol with the computer to receive the data and activate the transmission sequence of the data contained in the control console memory. The data can remain in the control console, or be permanently deleted. The green LED (Figure 3, item 23) of the control console (Figure 1, item 2) will be activated to confirm that the data transfer to the computer is being performed and the screen (Figure 3, item 21) will present a message with the state of data transfer.

Claims (12)

1. Morphometry system for the semi-automatic acquisition of three-dimensional coordinates of microscopic structures, especially suitable for Anatomy and Histology, with the purpose of acquiring simplified models of microscopic structures, characterized in that said system consists of two semi-rigid sensors ( 11, 12) and an opto-electronic sensor (13) connected to a control console (2) that stores the selected coordinates. 2. Morphometry system for the semi-automatic acquisition of three-dimensional coordinates, according to claim 1, characterized in that the two semi-rigid sensors (11, 12), are formed by two cylinders that are inserted into each other and inside which an analog linear potentiometer (115) is housed, whose cursor (112) is visible from the outside through a longitudinal opening in the outer cylinder and is adhered to the inner cylinder (113) which slides along the outer cylinder ( 114) producing a variable potential difference with the displacement that determines the lateral movement of the microscope plate (1). 3. Morphometry system for semi-automatic acquisition of three-dimensional coordinates, according to claim 2, so that in the two semi-rigid sensors (11, 12) the range of absolute displacement is characterized by two positions: the first is that in that the inner cylinder (113) is completely hidden inside the outer cylinder (114) and the second one is the one in which the inner cylinder (113) is completely outside the outer cylinder, so that its volume inside the outer cylinder (114) is minimal. 4. Morphometry system for the semi-automatic acquisition of three-dimensional coordinates, according to claims 2 and 3, characterized in that in the two semi-rigid sensors (11, 12) the distance between the minimum and the maximum position depends on the microscope and the resolution, which determine the positioning error caused by the analog transfer of the linear potentiometer. 5. Morphometry system for semi-automatic acquisition of three-dimensional coordinates, according to claim 1, characterized in that the opto-electronic sensor (13) used to measure the displacement of the microscope focus (1) is constituted by a socket (14) that it is coupled to the micrometric screw wheel of the focus and that at one of its bases a shaft with a flexible connection (131) is welded that allows transferring the rotation of the focus screw without loss to a grooved disc (133) on whose periphery have evenly distributed holes for the detection of the change of light to dark in the photosensor (132). 6. Morphometry system for the semi-automatic acquisition of three-dimensional coordinates, according to claims 1 and 5, characterized in that the opto-electronic sensor (13) is constituted by a photosensor (132) that detects the transition between light and dark produced by the slotted disc rotation (133) and by an opto-electronic counter (134) that determines the number of light-to-dark steps of each focus shift. 7. Morphometry system for the semi-automatic acquisition of three-dimensional coordinates, according to claims 1, 5 and 6, characterized in that the opto-electronic sensor (13) has a level of discretization appropriate to the problem and the micrometric screw wheel of the approach that Measures the vertical movement has the appropriate range for the required resolution. 8. Morphometry system for semi-automatic acquisition of three-dimensional coordinates, according to previous claims, characterized by a control console (2) that processes and stores the signals from the two semi-rigid sensors (11 and 12) and the sensor opto-electronic (13). 9. Morphometry system for the semi-automatic acquisition of three-dimensional coordinates, according to claim 8, characterized by a control console (2) formed by diodes indicating when the device is connected to the power grid and when data is being transferred from or towards the computer and the buttons sufficient for the selection of the work mode, so that the selection of the menu, the movements on the menu options, the cancellation of the current option, the confirmation of the information on the screen and the introduction of the coordinate recorded by the sensors for storage in the control console (2) is uniquely determined. 10. Morphometry system for semi-automatic acquisition of three-dimensional coordinates, according to claims 8 and 9, characterized by a control console (2) using a microcontroller managed by a program that controls the outputs of the three sensors (11, 12 and 13) and the inputs and options for the operation of the invention. 11. Morphometry system for the semi-automatic acquisition of three-dimensional coordinates, according to claim 10, characterized by using the digital inputs in the treatment of the keyboard signals, the liquid quartz screen, the signaling diodes and the sensor data input opto-electronic 12. Morphometry system for the semi-automatic acquisition of three-dimensional coordinates, according to claim 10, characterized by using the analog inputs for the two semi-rigid sensors that are connected to an analog-digital converter that allows to read directly the absolute value of the position of the X and Y displacement of the microscope plate.