Claims:CLAIMS
We claim:-

1. A modified organ bath adaptable to evaluate the effects of drugs and chemicals on the tissues, comprising of:-
an isolated tissue chamber (5) formed out of non-reactive cylindrical plastic;
a water bath (1) provides a stable temperature to an isolated tissue chamber (5) and a physiological salt solution tank (4) by the process of conduction of heat produced by heater (2);
a tissue holder (6) molded with the configuration of the isolated tissue chamber (5) where the tissue holds;
an electrical component compartment (10) providing oxygen to the tissue through an aeration pipe;
a digital imaging microscopic camera (7) having zooming capacity of 1000 times mechanically connected on the camera stand (9) with light (8) in the periphery;
wherein, the digital imaging microscopic camera (7) captures the image of the tissue being present in the isolated tissue chamber (5) and the image further processed through the artificial intelligence tools to detect the tissue area and pixel tracking.

2. The modified organ bath as claimed in claim 1 wherein, the isolated tissue chamber (5) comprising an outer chamber (51), a solution inlet pipe (52), the aeration inlet pipe (53) mechanically connected with the electrical component compartment (10), an inner perforated chamber (54), a solution outlet pipe (55) and a chamber lid (56). (numbering is illustrated)

3. The modified organ bath as claimed in claim 1 wherein, the tissue holder (6) further comprising elastic string (61) to provide tensile balance to the tissue and hook (62) as a point of attachment to the tissue.

4. The modified organ bath as claimed in claim 1 wherein, the physiological salt solution tank (4) and isolated chamber (5) mechanically connected with the motor through which the physiological salt solution from physiological salt solution tank (4) pumped into the isolated chamber (5) to provide nutrient to the tissue.

5. The modified organ bath as claimed in claim 1 wherein, the control unit (3) comprising the controlling knobs for aeration, temperature, pump inlet and outlet.

6. The modified organ bath as claimed in claim 1 wherein, chamber (1) comprising two main bodies, in which isolated tissue chamber (5) and physiological salt solution tank (4) and internally connected through the pump.

7. The modified organ bath as claimed in claim 1 wherein, an aeration inlet pipe (53) connected to the passage situated at the upper portion of outer chamber (51), and this aeration pipe (53) is wrapped in circular pattern at bottom of the chamber.

8. The modified organ bath as claimed in claim 1 wherein, the modified organ bath adaptable to evaluate the effects of drugs and chemicals on the tissues, wherein an isolated tissue chamber (5) formed out of non-reactive cylindrical plastic and a water bath (1) provides a stable temperature to an isolated chamber (5) and a physiological salt solution tank (4) by the process of conduction of heat produced by heater (2) in which a tissue holder (6) molded with the configuration of the isolated tissue chamber (5).

9. The modified organ bath as claimed in claim 1 wherein, electrical component compartment (10) provides oxygen to the tissue through the aeration inlet pipe (53).

Date this 20th June 2020 , Description:FORM 2
THE PATENTS ACT, 1970
(39 OF 1970)
&
The Patents Rules, 2003
COMPLETE SPECIFICATION
(See section 10; rule 13)
1. Title of the invention – A MODIFIED ORGAN BATH

2. Applicant
Name Nationality Address
IDRIS MITHAIWALA INDIAN Sakina manzil, Beside jamali apartment, Vohravad, Godhra, Gujarat
SUTHAR HETAL INDIAN 77/2 dhaval park,Kanjari road, Halol, Vadodara, Gujarat
JAMBUGHODAWALA SAKINA INDIAN Opp Session court, Vadodara highway, Godhra, Gujarat

3. PREAMBLE TO THE DESCRIPTION

The following specification particularly describes the invention and the manner in which it is to be performed.

FIELD OF THE INVENTION

The present invention relates to a modified organ bath. More particularly it relates with the organ bath which is having image processing tools which captures the images of tissue and process it through artificial intelligence system to provide accurate result in term of tissue response with any particular drugs.

BACKGROUND OF THE INVENTION

Biological assay is the in-vitro quantitative analytical technique, generally performed to estimate the potency and effectiveness of a substance by studying its effect on living cells or tissue. It is a comparative assessment of relative potency of a test compound to a standard compound. Understanding the potency of agonists and antagonists is a kind of bioassay usually performed using isolated organ bath.

Traditional organ bath, used in pharmacology laboratory and various educational institutes, is a combination of many small units and is utilized for investigating the effect of drugs on receptors in drug discovery in which the measurement of physiological tissue response to drug concentrations allows the generation of dose response curve. With this, the pharmacological profile of the drug can be determined by drug‘s EC50 i.e. concentration of drug required to obtain 50% effect and, IC50 i.e. measure of drug’s inhibition.

The set up of traditional organ bath includes the tissue/organ placed in organ tube filled with physiological salt solution and connected/tied through thread to properly balanced frontal writing lever by applying a load or tension in order to obtain the graphical amplified measurable response with the contraction and relaxation of tissue muscles through addition of respective drugs. This response is appeared on Kymograph paper glued on Sherrington recording drum. The organ tube containing tissues are maintained for several hours in a temperature controlled bath chamber and also supported with oxygen supply. The bath chamber can be of single unit and double unit which have one and two holding place for organ tube, respectively.

The slight improvement in the traditional organ bath is brought by the introduction of multichannel recordings due to the use of mechanical transducer, along with computer acquisition and analysis. Nevertheless, it also includes tying the tissue to lever using hook or thread and it may lead to the death of tissue due to tension generated and it also produces the inaccurate response sometime due to improper and uncontrolled aeration. Moreover, the transducers are not cost-effective which deters its frequent utilization.

There are many conventional modified and automated organ baths available in which the main advancement includes replacement of the any of the component of organ bath. But, there haven’t been much modifications as far as the integral component of the instrument is concerned i.e. tissue chamber. Furthermore, cleaning and assembling of each and every unit of multi-component instruments like conventional organ bath, while performing an experiment is a time consuming process and chances of error are more.

Further, an isolated tissue organ bath was working on mechanical based system that induces tensile force on tissue. Therefore many challenges arise while performing practical. The challenge is that to deal with the complex instrumental assembly setup, which is tedious. Which include balancing of writing lever rod, tying of tissue with thread, maintenance of tissue stability condition like temperature, aeration etc. The main challenge is to keep the tissue in its potent state.

Various prior art discloses about the organ bath. The prior art document 3283/DEL/2005 discloses the a tissue chamber made of non-reactive plastic and imprinted with graduation to fill the PSS solution as per requirement, further comprising water jacket surrounding the tissue chamber and containing water maintained at predetermined temperature with the help of thermostat. Here, the tissue holder has a dual role of holding the tissue and providing aeration. A transducer is incorporated to pick up the data relating to the major changes occurring due to contraction of tissue and converting the physiological data into electrical signal.

Another prior art document WO2007055705A1 disclosed a block for super fusing tissue with a fluid, comprising: a block body; a tissue chamber having a chamber volume with a defined capacity formed in said block body; an inflow passage in communication with said tissue chamber for providing the fluid to said tissue chamber; and an overflow passage in communication with said tissue chamber for receiving the fluid that exceeds said defined capacity of said chamber volume from said tissue chamber and directing the fluid away from said tissue chamber.

Another prior art document US4889691A disclosed a modular tissue super fusion chamber comprising: a magnetic base for supporting the chamber on a steel surface; support means; a support chamber rigidly secured to the support means, the support chamber having a single recess therein of a preselected size and geometry; temperature control means disposed in the support chamber and aligned with the recess for precisely controlling heat passing through the recess; and a plurality of modular bath containers for interchangeable mounting in the single recess, each having an identical external size and configuration matching that of the recess in the support chamber and each having a space therein of a different size and/or geometry for containing a bath and receiving tissue samples of different sizes and configurations, each modular container having ports there through for receiving at least one tube and at least one electrode, which electrode monitors electrophysiological characteristics of the tissue sample immersed in the bath.

The aforementioned prior arts encounter various problems which lead into the difficulties in many ways like a tissue holder with a hook which forms the reason for the strain of tissue and its ability of functioning as an aerator etc. Hence, it is desperately needed to introduce a modified organ bath which has features more efficient in comparison to existing one and also have additional features like direct graph printing facility, electrical tissue stimulation facility, accurate and rapid result obtained, no requirement of sherrington drum and many more.

OBJECT OF THE INVENTION

The principle object of the present invention is to provide a modified organ bath.

Another object of the present invention is to provide modified organ bath with stabilized temperature maintenance and unique arrangement of perforated tissue chamber which prohibits the fake responses generated due to air bubbles and provides excellent drug distribution.

Another object of the present invention is to provide modified organ bath which having bubble aeration control to avoid error.

Another object of the present invention is to provide modified organ bath provide auto temperature maintenance and no requirement of sherrington drum.

Another object of the present invention is to provide modified organ bath having facility of automatic tissue washing.

SUMMARY OF THE INVENTION

This summary is provided to introduce a selection of concepts in a simplified form that are further disclosed in the detailed description of the invention. This summary is not intended to identify key or essential inventive concepts of the claimed subject matter, nor is it intended for determining the scope of the claimed subject matter.

The present invention is all about a modified organ bath having image processing tools with the improved tissue chamber.

The main aspect of the present invention is to provide modified organ bath with stabilized temperature maintenance, unique arrangement of perforated tissue chamber which prohibits the fake responses generated due to air bubbles and provides excellent drug distribution and to generate the response use of digital imaging microscopic camera to capture the image of the tissue for the further image processing.

Another aspect of the present invention is to provide a modified organ bath adaptable to evaluate the effects of drugs and chemicals on the tissues, comprising of an isolated tissue chamber formed out of non-reactive cylindrical plastic; a water bath provides a stable temperature to an isolated chamber and a physiological salt solution tank by the process of conduction of heat produced by heater; a tissue holder molded with the configuration of the isolated tissue chamber where the tissue tying; an electrical component compartment providing oxygen to the tissue through an aeration pipe; a digital imaging microscopic camera having zooming capacity of 1000 times mechanically connected on the camera stand with light in the periphery; wherein, the digital imaging microscopic camera captures the image of the tissue being present in the isolated tissue chamber and the image further processed through the artificial intelligence tools to detect the tissue area and pixel tracking.

One more aspect of the present invention is to provide the isolated tissue chamber comprising an outer chamber, a solution inlet pipe, the aeration inlet pipe mechanically connected with the electrical component compartment, an inner perforated chamber, a solution outlet pipe and a chamber lid.

One another aspect of the present invention is that the tissue holder further comprising elastic string to provide tensile balance to the tissue and hook as a point of attachment to the tissue.

One another aspect of the present invention is that the physiological salt solution tank and isolated chamber mechanically connected with the motor through which the physiological salt solution from physiological salt solution tank pumped into the isolated chamber to provide nutrient to the tissue.

BRIEF DESCRIPTION OF DRAINGS

The foregoing summary, as well as the following detailed description of the invention, is better understood when read in conjunction with the appended drawings. For the purpose of illustrating the invention, exemplary constructions of the invention are shown in the drawings.

However, the invention is not limited to the specific methods and structures disclosed herein. The description of a method step or a structure referenced by a numeral in a drawing is applicable to the description of that method step or structure shown by that same numeral in any subsequent drawing herein.

Fig. 1 is a schematic presentation of the modified organ bath.

Fig. 2 is an illustrative representation of the isolated chamber and tissue holder.

Fig.3 is an experimental image of the tissue captured on regular time intervals.

Fig.4 is a graph of the (4a) doses graph Vs surface area and (4b) dose Vs response graph.

DETAILED DESCRIPTION OF INVENTION

The present invention overcomes the aforesaid drawbacks of the above and other objects, features and advantages of the present invention will now be described in greater detail. Also, the following description includes various specific details and is to be regarded as merely exemplary. Accordingly, those of ordinary skill in the art will recognize that: without departing from the scope and spirit of the present disclosure and its various embodiments there may be any number of changes and modifications described herein.

It must also be noted that as used herein and in the appended claims, the singular forms "a", "an," and "the" include plural references unless the context clearly dictates otherwise. Although any systems and methods similar or equivalent to those described herein can be used in the practice or testing of embodiments of the present invention, the preferred, systems and methods are now described.

"Optional" or "optionally' means that the subsequently described event or circumstance may or may not occur, and that the description includes instances where the event occurs and instances where it does not.

According to one embodiment, the present invention discloses a modified organ bath with improved tissue chamber.

The present invention is all about a modified organ bath having image processing tools with the improved tissue chamber.

An organ bath is used in pharmacology laboratory and various educational institutes, is utilized for investigating the effect of drugs on receptors in drug discovery in which the measurement of physiological tissue response to drug concentrations allows the generation of dose response curve.

The main embodiment of the present invention is to provide modified organ bath with stabilized temperature maintenance, unique arrangement of perforated tissue chamber which prohibits the fake responses generated due to air bubbles and provides excellent drug distribution and to generate the response use of digital imaging microscopic camera to capture the image of the tissue for the further image processing.

As per one embodiment of the present invention, the modified organ bath adaptable to evaluate the effects of drugs and chemicals on the tissues, comprising of an isolated tissue chamber (5) formed out of non-reactive cylindrical plastic; a water bath (1) provides a stable temperature to an isolated chamber (5) and a physiological salt solution tank (4) by the process of conduction of heat produced by heater (2); a tissue holder (6) molded with the configuration of the isolated tissue chamber (5) where the tissue tying; an electrical component compartment (10) providing oxygen to the tissue through an aeration pipe; a digital imaging microscopic camera (7) having zooming capacity of 1000 times mechanically connected on the camera stand (9) with light (8) in the periphery; wherein, the digital imaging microscopic camera (7) captures the image of the tissue being present in the isolated tissue chamber (5) and the image further processed through the artificial intelligence tools to detect the tissue area and pixel tracking.

As per another embodiment of the present invention the isolated tissue chamber (5) comprising an outer chamber (51), a solution inlet pipe (52), the aeration inlet pipe (53) mechanically connected with the electrical component compartment (10), an inner perforated chamber (54), a solution outlet pipe (55) and a chamber lid (56).

As per another embodiment of the present invention the physiologic salt solution is a 0.9 % solution of sodium chloride and water; it is isotonic, i.e., of the same osmotic pressure as blood serum. The physiological salt solution is isotonic with body fluids, it may also be used as a solvent or diluents, for antibiotics and other pharmaceuticals and biological where compatible, and for washing mucous membranes and other tissue surfaces.

As per another embodiment of the present invention the electrical component compartment (10) provides oxygen to the tissue through the aeration inlet pipe (53). While the chamber is filled with the solution and the switch on the aerator, it produces bubbles which can disturb the camera and can also cause tissue movement. To avoid this situation, the inner perforated chamber (54) mounted in the isolated chamber. The gas bubbles will hit on the surface of the inner perforated chamber (53) and release the oxygen which than provided to the tissue in the chamber.

In the following, a first embodiment of the present invention will be described with reference to Fig. 1.

According to it, the modified water bath chamber (1) comprising two main bodies i.e. isolated tissue chamber (5) and physiological salt solution tank (4) and internally connected through the pump. The isolated tissue chamber contains the tissue holder (6) for holding the tissue. To provide the heat to the solution filled in the physiological salt solution tank (4) the heater (2) being molded below. For the purpose of the image processing tools the digital imaging microscopic camera (7) with light (8) in the periphery molded on the camera stand (9) as the camera can easily capture the image of the tissue. For getting the better response from the tissue the oxygen provided from the electrical component compartment (10) through the aeration pipe. As per the controlling element the controlling unit (3) comprising the controlling knobs for aeration, temperature, pump inlet and outlet.

In the following, the illustrated embodiment of the present invention will be described in Fig. 2.

According to it, the tissue chamber is double concentric circle shaped chamber. There is an aeration inlet pipe (53) connected to the passage situated at the upper portion of outer chamber (51), and this aeration pipe (53) is wrapped in circular pattern at bottom of the chamber. There are holes on the top of the chamber with a small fan like structure which rotates with the force of air. When the organ bath switch is in the operation mode than the solution from the physiological salt solution tank (4) will transfer through the solution inlet pipe (52). To avoid the error cause by the bubble the inner perforated chamber (54) situated in the isolated tissue chamber (5). For removing the solution from the chamber the solution outlet pipe (55) situated at the bottom of the chamber. To avoid unnecessary effect the physiological salt solution tank (4) covered through the chamber lid (56).

The present invention was experimented and illustrated more in details in the following example. The example describes and demonstrates embodiments within the scope of the present invention. This example was given solely for the purpose of illustration and is not to be construed as limitations of the present invention, as many variations thereof are possible without departing from spirit and scope.

While performing the experiment, filling the solution and placing of the tissue chamber are done in the respective compartments. The tissue stand having a tissue is placed upon the tissue chamber, apparatus is switched on, and PSS solution starts moving to the chamber compartment which makes the inner chamber filled. With the aerator functioning, the bubbles from the passage of outer chamber generate the force through which fan starts moving in circular manner. Furthermore, the bubbles coming from outer chamber are blocked by the perforations of the inner chamber, and this will produce air inside the inner chamber. Due to the circular movement of fan, when the drug is injected in the inner chamber containing tissue, it is distributed equally. The response produced by the tissue is captured by the digital imaging microscopic camera fixed upon the tissue chamber compartment and directed to the artificial intelligence tools. The tools process the image and generate the graph of pixel v/s time.

Example 1: The experiments and the result carried out using Image sensitize modified organ bath is as follows-

Aim: Endpoint bioassay using Acetylcholine on chicken ileum.
Requirements: Chicken ileum
Chemicals: All other chemicals used are of laboratory grade. Sodium Chloride, Potassium Chloride, Calcium Chloride, Magnesium Chloride, Magnesium Sulphate, Sodium bicarbonate, Sodium hydrogen phosphate, Potassium hydrogen phosphate and distilled water. Acetylcholine stock solution (1mg/ml)

Preparation of physiological salt solution (PSS):-
Compound Frog Ringer Ringer Locke De Jalon Tyrode Krebs
Nacl 6 9.0 9 8.0 6.9
Kcl 0.14 0.42 0.42 0.2 0.35
CaCl2 0.12 0.24 0.06 0.2 0.28
MgCl2 - - - 0.10 -
MgSo4 - - - - 1.28
NaHco3 0.2 0.5 0.5 1.0 2.1
NaH2Po4 - - - 0.05 -
KH2Po4 - - - - 0.16
Glucose 2 1.0 0.5 1 or 2 1 or 2

Method:-

1. Fresh entire gastrointestinal tract of healthy cock was obtained from a slaughter house.
2. The caecum was lifted forwards and the ileocaecal junction was identified.
3. A few centimetres of the ileum portion was cut and removed and immediately placed it in the watch glass containing physiological salt solution. The mesentery and adhering tissues were removed with gentle care.
Utmost care was taken to avoid any damage to the gut muscle. The ileum was cut into small segments of 2-3 cm long.
4. The piece of ileum was measured around 2cm accurately and was mounted on the tissue holder (6) with elastic string (61) and hook (62) at an angle of 1800. The tissue was then immersed into isolated tissue chamber (5) containing PSS maintained at 32-35°C and bubbled with air.
5. The system was switched on and the tissue was adjusted with the lighting condition upon proper selection of each part of tissue. These images is shown in Fig. 3.
6. The response was taken and at different time interval upon addition of doses the time and the dose was noted.
7. After completion of the experiment the graph of surface area vs time is compared with dose and Dose vs Response table is prepared through the artificial intelligence. These graphs are shown in Fig 4.
8. Calculate Dose vs Response by using following formula
Response= Minimum contraction/Maximum contraction*100

Conclusion:
1. ISM Organ bath with a future scope replacing the current one with very low cost available equipment which can be implemented in any academic or industrial laboratories.
2. Automatic technologies provide less interface of the user with more accuracy and facilities to work on and provide efficient results
3. The product aims to provide more accurate and rapid development of the drug research in field of animal model based drug evaluation with a greater, convenient and automated amenity.
4. The product available at a cheaper cost, has a drastic impact on economic factor ensuring each educational and research lab can afford it.
5. Minimizing the tissue usage for the experiment tends to save the number of animal sacrificed during the experimentation

REFERENCE NUMERALS
Water bath (1)
Heater (2)
Control unit (3)
Physiological salt solution tank (4)
Isolated tissue chamber (5)
Outer chamber (51)
Solution inlet pipe (52)
Aeration inlet pipe (53)
Inner perforated chamber (54)
Solution outlet pipe (55)
Chamber lid (56)
Tissue holder (6)
Elastic string (61)
Hook (62)
Digital imaging microscopic camera (7)
Light (8)
Camera stand (9)
Electrical component compartment (10)