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DIAGNOSTIC APPLICATION OF
DARKFIELD MICROSCOPY
 
Author: Stephen C. Tamplin
RN ND DAcup DCHyp PhD(Alt Med)
 
 
Abstract, and extract from thesis.
 
This thesis was undertaken as a research project leading to the award of the degree of Doctor of Philosophy in 2012.
 
Darkfield microscopy is a process whereby peripheral blood samples can be observed in the clinical setting. It is not a process belonging exclusively in the realm of medicine, as it is used by health care practitioners in both conventional western medicine and alternative (or natural) medicines, veterinary work, and in non health-care fields such as geology, where gem-stone clarity is validated.
 
This study explored the history of microscopy, and examined the relationship between darkfield microscopy, naturopathic diagnosis and haematology, and demonstrated the further use and scientific application of darkfield microscopy in the clinical setting.
 
Comparison with the ‘smear and stain’ technique used in brightfield microscopy and automated cell counting was undertaken.
 
A review of practitioner education, microscopy techniques and interpretation of observations was part of the research project.
 
Although it is a process used as a screening tool for live blood cell analysis for over 100 years its outcomes have not been generally accepted as reliable or valid by western medical pathology.
 
This thesis does not seek to discuss all blood parameters observed by darkfield microscopy, but to demonstrate the application of darkfield microscopy as a point of care diagnostic device.
 

Research question:
 
Is darkfield microscopy able to provide a consistent, reliable and valid outcome supporting its use as a clinical diagnostic tool?
 
Hypothesis:
Using known haematological parameters, darkfield microscopy observations can be shown to provide sufficient reliability to allow its use in either naturopathic or haematological fields to make or contribute to authorative clinical diagnosis.
 
INTRODUCTION, AND REASON FOR THE STUDY.
This subject was chosen for research due to the many claims and counter claims made by various ‘experts’, from both naturopathic and medical fields, and dispute between practitioners and pathologists on what are or are not observed pathologies or artifacts within or upon specimens, which have cast doubt on the reliability of darkfield microscopy as a valid analytical procedure.
 
There is much contradiction of the reliability, validity and interpretation of observations, and confusion about the prescriptive path required from those observations by naturopathic practitioners that needs to be clarified.
 
Within the natural medicine / alternative medicine fraternity there are practitioners using darkfield microscopy who have minimal haematological knowledge, some who rigidly adhere to the work of Enderlein with disregard to contemporary scientific knowledge, others who have made interpretations of darkfield microscopy an art-form of myth and ‘parasitism’ as their basis of treatment, while others are incorrect in their observation due to a lack of basic understanding of the body’s structure and function at even a macro level, and are unable to conduct a thorough and professional clinical examination.
 
Further, discrepancies have been noted in methods used for slide preparation, peripheral blood sampling and the relationship of darkfield microscopy to the results of basic urine biochemistry and other blood pathology testing, such as by smear and stain technique, and automated measurement of blood factors. 
 
The relationship between cellular nutrition and healthy organ function is also a matter of biological versus medical science. Darkfield microscopy provides a biological understanding of health disturbances, although some in the ‘medical science’ field discredit this basis.
 
By using a nutritional approach to case management, observations may be made which support darkfield microscopy as a useful point-of-care diagnostic tool for wholistic health care. It is shown how in main stream medicine darkfield microscopy is a validated tool, described as a ‘gold standard’ for some diagnostic requirements.
 
A metasearch of literature and general web-searches showed varying opinion (both informed and arrogant) among commentators, opinion which may be biased according to the commentator’s primary field of training, and knowledge or lack thereof of basic hematological science, understanding of darkfield parameters, and nutritional causes of disturbance of health.  Blog sites abound with ridicule and defense of the procedure of live blood analysis, with no consensus on how interpretations of the procedure are made.
 
Metasearch engines Ixquick.com, MonsterCrawler.com and Google.com reveal very few actual research papers on darkfield microscopy appear to exist.  
 
There is research which admits use of darkfield microscopy in its process, but not of the darkfield process itself.
 
Most of the web-sites presented by meta-search are advertisements for darkfield equipment (from suppliers) or numerous ‘cut and paste’ articles from clinics and practitioners promoting darkfield microscopy for live blood cell analysis.
 
The nature of the information presented in these articles has a common theme and similar presentation, suggesting the web-site host has not undertaken individual research, but possibly personalised (plagiarized) articles presented by some other source.
 
THE RESEARCH COVERED THE FOLLOWING TOPICS:
 
1.           Training and education of practitioners
2.           Slide preparation
3.           Blood sample extraction
4.           Technique validity and time-delayed observations
5.           Venous vv peripheral blood
6.           Fasting vv non-fasting samples
7.             Comparison with other point of care diagnostic devices
8.             Non cellular components of plasma
9.             Oxidative stress and disease process
10.        Darkfield microscopy and cancer
 
As well a comparison of haematological morphology with Darkfield observations and Enderlein’s descriptions is tabled.
 
OBSERVATIONS OF THE RESEARCH OUTCOMES
The research tasks demonstrated that deficiencies exist in training of, and current technique used by, darkfield microscopists. The awareness of these deficiencies supported doubts on the reliability of darkfield microscopy to provide a consistent, reliable and valid outcome.
 
Through the comparative analysis of haematological parameters and darkfield observations there is sufficient consistency between the observations made by ‘smear and stain’ slide and darkfield slide to enable reliability of the technique, in the clinical setting.
 
Artifact errors have been shown to contribute to gross error both in clinical assumption and published work. This can lead to ridicule or rejection of observations and discredit darkfield microscopy in general.
 
By adopting a standardised ‘best practice’ for slide preparation and ensuring a measured or consistent quantity of peripheral blood sample the reliability can be further improved.
 
The respondents to the survey indicated a need for improved training, at a more recognised, formal level. This could be achieved by placing darkfield microscopy in a medical laboratory technology program, using recognised haematological nomenclature and technique, with exposure to brightfield processes. Naming of observations using correct haematological identifiers as well as a thorough understanding of nutrition associated morphology is critical to broader acceptance of this technology.
 
The science of haematology is profound, and natural medicine practitioners are not generally skilled in interpretation of that science, however darkfield microscopy provides clinicians with evidence supporting nutritional relevance in the biological sciences, and health education.
 
In contrast medical practitioners are not well trained in nutrition, and darkfield microscopy is an invaluable tool for educational outcomes, as it links biologically the fields of nutrition and haematology.
 
That darkfield microscopy is taught in some medical schools, and is the gold-standard for certain parasitic diagnostics lends weight to the argument for the hypothesis, that reliable outcomes can be achieved with this technique.
 
Using a darkfield microscope as a point-of-care diagnostic device, clinicians can undertake an essential part of a reliable triple test upon which a diagnosis can be based.
 
ANALYSIS OF DATA.
One of the key problems with the thesis was been the unwillingness of other darkfield microscopy practitioners to be involved in this research project.
 
A cohort group of 20 practitioners were approached to participate in aspects of this research project. It became apparent early in the research that practitioners were in general reluctant to participate in this research, citing lack of confidence and unwillingness to have their interpretations peer-reviewed or subject to scrutiny.
 
In order to professionalise the practice of darkfield microscopy, increasing practitioner confidence, training should be formalized under a system similar to the Australian Vocational Education certification levels, at least at an associate diploma level, such as an Associate Diploma of Darkfield Laboratory Technology, ie a  level 5 qualification, with learning taking place over more than just a few days. In Australia level 6 is bachelor degree level, which requires a four year period of study.
 
A Vocational Education Training Package could be developed in conjunction with haematology, medical laboratory and wholistic practitioners to standardise the nomenclature, practice and interpretation of darkfield observations.
 
By using a standardised approach to sample procurement and slide preparation procedural and artifact errors would be reduced, thus improving the accuracy of interpretation and enabling repeatable results, one of the desired tenets of scientific scrutiny. The research identifies a simple best practice technique which in clinical practice is not onerous or time consuming, but does provide improved outcomes.
 
The work, either partly or in whole, and the conclusions drawn may be open to question, even dispute, however those questions or disputes are best answered by scientific study of a larger sample. The main outcome of this study has been to clarify best practice, from which standardised processes may be defined to improve the status of the procedure.
 
Alignment of the process with recognised haematological pathophysiology has been demonstrated, and the biological / nutritional impact on haematology has been recognised. The parallels between haematology and peripheral blood analysis by darkfield microscopy are identified as being more closer than distant, and the benefits from use of darkfield microscopy in the clinical setting as a point-of-care diagnostic tool have been proven.
 
The research demonstrated that while a cursory 10 to 15 minute observation of the slide provides an insight to a subject’s health state, it is an inadequate opportunity to identify clinically significant processes, as the significant changes observed on slides in this research did not always occur within the suggested time.
 
Nutritional and degenerative organ-related causes of sickness and disease can be observed in the early part of observations however additional time to identify pathological causes, such as fungal or bacterial processes needs to be properly allowed for.
 
As the rate of change in the sample is related to the nature of the client’s condition, if there is suspicion of chronic or obscure disease states longer periods of observation will be required.
 
The gathered data, both numeric and pictorial provide graphic evidence of the value of darkfield microscopy, and validate its diagnostic use.
 
At the outset, this work asked the question:
 
“Is darkfield microscopy able to provide a consistent, reliable and valid outcome supporting its use as a clinical diagnostic tool?”
 
This author believes his study of the process and the outcomes presented answer in the affirmative, but there are areas in which the process can be improved.
 
Webinars are being arranged to inform interested practitioners and students of the findings of the thesis, and these will be published in industry journals and supplier newsletters.
 
 

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